western blotting brochure

Western Blotting

Some protein detection kitsmake you wonder if your eyesare playing tricks on you.

SEE MORE with KPL!

Western Blotting

KPL’s blotting substrates and Protein

Detector™ Western Blot Kits pro-

vide consistent, high quality results

when analyzing proteins immobi-

lized on membrane. Choose from a

line of substrates and kits for either

chemiluminescent or colorimetric

methods of detecting specific anti-

gens in a complex protein mixture.

The cornerstone of KPL’s Western

blotting systems is our line of sensi-

tive liquid substrates. You have a

choice among multiple chemilumi-

nescent and chromogenic substrates

for peroxidase and phosphatase that

provide varied levels of sensitivity.

The chemiluminescent substrates

provide a reliable alternative to con-

ventional colorimetric ELISA with

the advantage of increased sensitivity.

Our chemiluminescent substrates

for alkaline phosphatase detection,

PhosphaGLO™ and PhosphaGLO

Reserve™ AP Chemiluminescent

Substrates, offer sensitivity and

duration of signal in a one-compo-

nent system.

KPL’s substrates are offered as stand-

alone reagents or combined with

unique blocking and wash solutions

and highly specific secondary anti-

bodies to create comprehensive,

fully optimized kits for Western

blotting. Each of these kits is

designed to deliver the maximum

signal-to-noise ratio available.

Choose the detection system that

best meets your needs and achieve

clean, clear results blot after blot.

No matter which KPL Western Blotting system you use,you’ll get a strong, clean signal every time.

Substrates for Western Blotting

KPL’s Western blotting substrates for use in peroxidase and phosphatase reporter systems offer high quality, reproduciblechemiluminescent and colorimetric detection. It’s your choice!

comp = component • chemi. = chemiluminescent • color. = colorimetric • HRP = peroxidase • AP = phosphatase • NA = not applicable

LumiGLO LumiGLO PhosphaGLO PhosphaGLO TMB HRP 4 CN BCIP/NBT FirePhosReserve HRP HRP Reserve AP AP Substrate Substrate Substrate AP Substrate MembraneSubstrate Substrate Substrate AP Substrate

Type chemi. chemi. chemi. chemi. color. color. color. color.

Format 2-comp 2-comp 1-comp 1-comp 1- or 3- 2-comp 1- or 3- 1-compcomp comp

Detection film or image film or image film or image film or image visual-dark blue visual - purple visual - purple visual - redMethod analysis analysis analysis analysis precipitate precipitate precipitate

Detection Sub-picogram; 0.6 picogram femtogram picogram 50 picogram 500 picogram low nanogram low nanogramLimit femtogram

Stability of 8 hours 24 hours 2 years 2 years 1-comp- 1 hour 1-comp- NAWorking Solution at 4o C at 4o C 1 year 1 year

at 4o C. at 4o C.3-comp- 3-comp- 24 hours 1 hour

Duration of 4 - 8 hours 1 - 2 hours 5 days 5 days NA NA NA NASignal

Enzyme HRP HRP AP AP HRP HRP AP APCatalyst

Kinetics 5 minutes 5 minutes 15 minutes 15 minutes 10 minutes 10 minutes 10 minutes 10 minutes

Recommended nitrocellulose nitrocellulose nitrocellulose nitrocellulose nitrocellulose nitrocellulose nitrocellulose nitrocelluloseMembrane or PVDF or PVDF or PVDF or PVDF or PVDF or PVDF or PVDF or PVDF

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With KPL’s Protein Detector™

Western Blotting Systems, theresults jump right out at you.

Rapid, Sensitive Detection withLuminol-based SubstratesChemiluminescent detection involves

the creation of light through the catalysis

of an enzyme substrate. Use of this

method for protein detection allows an

increase in sensitivity by orders of mag-

nitude compared to traditional colori-

metric Western or dot blotting. KPL

offers two unique luminol-based chemi-

luminescent substrates —LumiGLO®

and LumiGLO Reserve™, for the rapid

and sensitive detection of horseradish

peroxidase (HRP)-labeled conjugates.

Maximum Sensitivity withLumiGLO ReserveTM

LumiGLO Reserve HRP Chemilum-

inescent Substrate offers an option for

those assays where enhanced sensitivity

is critical to success. This proprietary

two-component substrate formulation

provides greater than 20 times the sensi-

tivity of standard LumiGLO with detec-

tion levels as low as the femtogram

range (Figure 1).

Long Signal DurationLumiGLO Reserve emits light over the

course of 4-8 hours with the most

intense emission occurring within the

first two hours. Because of its extreme

light intensity, most images may be cap-

tured well under 10 minutes; multiple

exposures are easy to obtain. High sig-

nal intensity facilitates the detection of

both high and low abundance proteins

and makes it an ideal system for use

with chemiluminescent imagers.

Sample and AntibodyConservationLumiGLO Reserve provides the added

benefit of strong signal with the use of

reduced amounts of precious target and

antibodies. Therefore, material of limit-

ed supply or higher expense can be con-

served while maintaining your current

level of sensitivity.

Superior Signal to NoiseLumiGLO Reserve delivers lower non-

specific signal than competitor sub-

strates in its class (Figures 2 and 3).

ConvenienceLumiGLO Reserve Chemiluminescent

Substrate Kits are supplied in several kit

formats to meet diverse user needs.

This substrate can replace your current

chemiluminescent substrate in existing

assays where greater sensitivity is desired.

LumiGLO ReserveWestern Blot KitLumiGLO Reserve is also offered as part

of a fully optimized kit, the Protein

Detector LumiGLO Reserve Western Blot

Kit, providing femtogram detection of

proteins immobilized on membranes.

For assurance against background inter-

ference, this kit contains our unique

Detector™ Block for optimal signal-to-

noise ratio; very low background can be

achieved without compromise to signal

intensity (Figures 2 and 3). Detector

Block is also offered separately.

Reliable and EconomicalLumiGLO®

KPL’s original LumiGLO Chemilum-

inescent Substrate provides high quality

results in a variety of immuoassays.

Chemiluminescent Substrates and Western Blot Kits

Figure 1: Relative expression of transcription factor,c-myc, using different chemiluminescent substrates. Fivetwo-fold serial dilutions of purified c-myc (25 ng–1.56ng, lanes 1–5) were compared to a 64 µg total proteinHeLa nuclear lysate (lane 6). Following separation on a4-20% PAGE gel and transfer to PVDF, protein wasdetected using a rabbit anti-c-myc antibody (1:200)and anti-rabbit HRP conjugate (1:10,000). Detectionconditions were identical with the exception of substrate.

While the c-myc lysate sample was not detectable withA) LumiGLO or C) ECL Plus™ after 10 minutes, thesample was easily detected with B) LumiGLO Reserveafter just a 2-minute film exposure.

1 2 3 4 5 6

Figure 2: Comparison of low-end sensitivity using LumiGLO Reserve and ECLDetection Kits. Two-fold serial dilutions of Mouse IgG (1 ng – 31 pg) wereseparated by SDS-PAGE and transferred to PVDF. Under manufacturer’s recommend-ed conditions, protein was detected using HRP-labeled anti-mouse antibody (varieddilutions according to recommended optimization) and each respective substrate:A) LumiGLO Reserve, B) ECL Plus™, C) ECL Advance™. Film was exposed for 10minutes and analyzed for sensitivity and signal to noise.

A B C

Western Blotting

LumiGLO Reserve Signal-to-Noise Comparison

LumiGLO Reserve ECL Plus

LumiGLO

LumiGLO Reserve

ECL Plus

ECL Advance

A

B

C

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Figure 5: Detection of ß-galactosidase in Western blots using alternativechemiluminescent substrates. A two-fold dilution series of purifiedß-galactosidase (from 25 was electrophoresed on a polyacrylamide geland transferred to KODAK BIOMAX Multi-Blot Kit for Proteins. Each blotwas detected under the same conditions using Protein Detector LumiGLOWestern Blotting Kit, substituting 5 mL LumiGLO on 3 of the4 blots with 5 mL of leading competitive chemiluminescent substrates,respectively. Following a 10 minute film exposure, results were evaluatedfor sensitivity and signal:noise ratio.

ECL™

Supersignal® West Pico

ECL Plus™

LumiGLO ChemiluminescentPeroxidase Substrate

Figure 4: Signal response comparison of LumiGLO vs. ECL™chemiluminescent substrates in Western blotting. ß-galactosidase waselectrophoresed, transferred to PVDF membrane and subsequentlydetected using rabbit anti-ß-gal followed by HRP conjugated goatanti-rabbit IgG (H+L). Each blot was treated with 5 mL of LumiGLO orECL substrate and exposed to film for 10 minutes. The density of eachband was analyzed on a Syngene GeneGenius™ image analyzer, usingautomatic background subtract.

LumiGLO vs. Leading Traditional Chemiluminescent Substrates

Figure 3: Signal linearity and signal-to-noise ratio comparison of advanced chemiluminescent substrates.Mouse IgG (250 pg to 3.91 pg) was transferred to nitrocellulose. Blots were detected under identical condi-tions with a 1:10,000 dilution of HRP-Goat-anti-Mouse IgG (except ECL Advance, 1:100,000) and exposed tofilm for 10 minutes. Densitometry was then performed using the Syngene GeneGenius. LumiGLO Reservedemonstrates superior signal linearity over a larger dynamic range and the greatest comparative signal-to-noise.

Superior Signal to Noise with Detector™ Block

Researchers continue to select

LumiGLO for reliable performance in

Western blot detection.

Ideal for Routine Western Blotting

LumiGLO detects low picogram quanti-

ties of target protein on blots. After

reaction with membrane-bound HRP

conjugates, light emission begins imme-

diately and continues for approximately

1–2 hours. Detection is possible within

minutes of blot exposure.

Greater Linearity

LumiGLO produces a quantitatively lin-

ear signal on film that ensures a broad-

er dynamic range of detection. While

the light output from other chemilumi-

nescent substrates tends to reduce

sharply as the concentration of protein

is titrated, a proportional reduction in

sensitivity is achieved with LumiGLO

(Figure 4).

Low Background

LumiGLO delivers superior signal-to-

noise by design. When used with KPL’s

Detector™ Block, nonspecific binding is

further reduced and ensures low back-

ground without loss of signal intensity.

Figure 5 compares the superior signal-

to-noise from blots blocked with

Detector Block and subsequently

detected with LumiGLO to leading

competitor substrates.

Cost Effective

LumiGLO is more economical blot for

blot compared to competitive products.

Enjoy the benefits of a sensitive,

reliable substrate while staying within

budget.

LumiGLO Western Blot Kit

The LumiGLO Western Blot Kit is

designed for low picogram detection of

proteins immobilized on membranes.

Researchers choose this kit for its reli-

able, consistent performance. The com-

bination of a stable, liquid conjugate

and a sensitive chemiluminescent

substrate allows rapid and accurate

identification of samples. For assurance

against background interference, this

kit contains our unique Detector Block

for optimal signal-to- noise ratio

(Figure 5).

SEE MORE with KPL!

0

Densitometry for LumiGLO vs. ECL

Rel

ativ

eIm

age

Den

sity

Concentration (ng)

45000

50000

40000

35000

30000

25000

20000

15000

10000

5000

0 50 100 150 200 250

LumiGLO

ECL

Sensitivity and Convenience withPhosphaGLOTM AP Substrates

KPL offers two chemiluminescent sub-

strates for use with alkaline phosphatase,

PhosphaGLO ReserveTM and PhosphaGLOTM

AP Substrates. PhosphaGLO Reserve

Substrate overcomes the limitations

posed by conventional chemiluminescent

substrates for AP. With sensitivity in the

femtogram range, PhosphaGLO Reserve

enables detection of target protein in low

concentrations. PhosphaGLO AP

Substrate is recommended for routine

detection of proteins in the picogram

range (Figure 6).

Low Background without

Special Blockers

PhosphaGLO Substrates produce superi-

or signal and low background, providing

a better signal to noise ratio and a cleaner

image than other chemiluminescent sub-

strates for AP (Figure 6). No special

blockers are needed with either nitrocel-

lulose or PVDF membrane.

Ultimate Convenience

Both PhosphaGLO AP Substrates offer

exceptional convenience as one-compo-

nent solutions ready to use. They are

stable for up to two years when stored at

4o C. Their long glow times of 5 days

facilitate assay development, enabling

repeat exposures.

KPL also offers colorimetric Western blot

systems when chemiluminescent detec-

tion is not preferred. Results can be inter-

preted by direct visualization of the blot.

Everything you need to detect your anti-

gen and primary antibody is supplied for

convenience and optimal performance.

Protein DetectorTM TMBWestern Blot Kit

The Protein DetectorTM TMB Western Blot

Kit utilizes TMB Peroxidase Membrane

Substrate (3,3’,5,5’-tetramethylbenzi-

dine), the most sensitive chromogenic

peroxidase substrate for Western and dot

blotting applications. Detection limits are

significantly increased as compared to

other chromogenic membrane

substrates. TMB produces a dark blue

precipitate upon reaction with HRP.

Protein DetectorTM BCIP/NBTWestern Blot Kit

For detection of phosphatase-labeled

conjugates, the Protein Detector

BCIP/NBT Western Blot Kit is ideal.

When reacted with alkaline phosphatase,

BCIP/NBT produces clean, intense bands

of purple precipitate. It also provides

stable, more permanent results than

other chromogenic substrates.

Western Blotting

In addition to our line of substrates and kits,

KPL offers a broad line of secondary antibodies,

conjugates and support reagents in various

packaging formats to suit your needs.

Secondary Antibodies

• Highly purified and specific.

• Large offering of species-specific polyclonal

antibodies

• Enzyme, biotin and fluorochrome-labeled

antibodies

Support Reagents

• One source for block solutions, wash

buffers, diluents and stabilizers.

• Consistently produced for reliable results.

LumiGLO is a registered trademark and LumiGLOReserve, Protein Detector, PhosphaGLO, PhosphaGLOReserve, FirePhos and Detector are trademarksof KPL, Inc.ECL, ECL Plus and ECL Advance are trademarks of GEHealthcare.SuperSignal West is a registered trademark of PierceBiotechnology, Inc.BIOMAX is a trademark of KODAK.GeneGenius is a trademark of Syngene.Biodyne is a registered trademark of Pall/GelmanCorporation.CDP-Star is a registered trademark of AppliedBiosystems.

Figure 6: Comparison of low-end sensitivity using PhosphaGLO Reserve AP Substrate, PhosphaGLO APSubstrate, and CDP-Star Chemiluminescent Substrate on PVDF membranes. Five-fold serial dilutions ofmouse IgG (2 ng -3.2 pg) were separated by SDS-PAGE and transferred to membranes. Protein wasdetected using a 1:1000 dilution of biotin-labeled goat anti-mouse IgG, a 1:10,000 dilution of AP-labeledstreptavidin and each respective substrate. Film was exposed for 10 minutes.

Colorimetric Kits

PhosphaGLO Reserve PhosphaGLO CDP Star®

AP Substrate AP Substrate ChemiluminescentSubstrate

Western Blot Reagents

SEE MORE with KPL!

BCIP/NBT (5-Bromo-4-chloro-3-indolyl phosphate/

nitroblue tetrazolium) is the most commonly used

substrate for alkaline phosphatase (AP) detection in

Western blotting due to its high sensitivity and low

background staining. Traditional formulations of

BCIP/NBT produce a deep purple color when reacted

with AP conjugates.

Now KPL’s new BCIP/INT formulation, FirePhosTM

AP Membrane Substrate, forms a red precipitate, pro-

viding a choice of red or purple precipitating AP sub-

strates to better optimize your individual assays.

FirePhosTM AP Membrane Substrate

BCIP/NBT Phosphatase Substrate

Sensitive and reproducible

No matter which substrate you choose, you’ll achieve

sensitive and reproducible detection of proteins on

Western blots. Both substrates provide sensitivity in

the low nanogram range with minimal background

(Figure 1) , sharp band resolution and a clear image.

The color produced is stable and linear over a wide

dynamic range.

See RED and PURPLE in your Western blots!

SEE MORE with KPL!

AP Membrane Color Substrates

FirePhos AP BCIP/NBT Substrate BCIP/NBT SubstrateMembrane Substrate (1-Component) (3-Component)

Figure 1: KPL’s substrates for alkaline phosphatasedetection in Western blotting demonstrate high sen-sitivity and low background. A two-fold dilutionseries from 250 ng to 3.9 ng of mouse IgG was loadedonto a Tris-HCl gradient gel, then blotted onto nitro-cellulose. The antigen was detected with ReserveAPTM

Goat Anti-mouse Conjugate (KPL) followed byFirePhos AP Membrane and BCIP/NBT Substrates (1-component and 3-component).

Ordering Information

To order or for more information, contact us at 800.638.3167 / 301.948.7755,

fax 301.948.0169 or visit us at www.kpl.com.

FirePhos and ReserveAP are trademarks of KPL.

Catalog# Description Size

FirePhosTM AP Membrane Substrate

50-81-30 FirePhos Membrane Phosphatase Substrate (1-Component) 100 mL

50-81-40 FirePhos Membrane Phosphatase Substrate (1-Component) 400 mL

50-81-34 FirePhos Membrane Phosphatase Substratee (1-Component) 1 L

BCIP/NBT Phosphatase Membrane Substrate

50-81-18 BCIP/NBT Membrane Phosphatase Substrate (1-Component) 100 mL


50-81-10 BCIP/NBT Membrane Phosphatase Substrate (1-Component) 1 L


Membrane Blocking

71-83-00 Detector Block (5X) 240 mL

KPL’s colorimetric membrane substrates are part of our comprehensive line of Western blottng

kits and reagents. Ask about our AP chemiluminescent substrates that can significantly lower

the detection limit of an assay.

ReserveAPTM

Conjugates

Gaithersburg, MD 20878

Phone: 800.638.3167

Fax: 301.948.0169

www.kpl.com

ISO 9001:2000 Registered

ML348-02

For research use only.© 2007 KPL, Inc. All rights reserved.

Improve your assay performance even furtherwith ReserveAPTM-labeledAntibodies!

KPL’s new ReserveAPTM Alkaline

Phosphatase-labeled Conjugates

exhibit high potency and consistent

performance in immunoassays.

These conjugates are the result of

advances in our conjugation chem-

istry and offer high signal and low

background while meeting KPL’s

standards for stability and repro-

ducibility. They are ideal for

demanding immunoassays that

require high detection sensitivity,

including ELISA, Western blotting

and immunohistology. Visit

www.kpl.com for more information

on KPL’s ReserveAP conjugates.

SEE MORE with KPL!

SEE MORE with KPL!

Non-fading

FirePhos and BCIP/NBT produce a bright red or purple precipitate that can be

deposited on nitrocellulose and PVDF membranes. They resist fading when

exposed to light.

Convenient

These substrates are supplied as convenient one-component solutions that are

ready to use. KPL’s BCIP/NBT is also available as a three-component solution.

Both substrates are provided in several sizes to meet your needs.

AP Membrane Color Substrates

KPL offers two substrates commonly used to detect

horseradish peroxidase (HRP) conjugated antibodies

that bring complementary benefits to membrane

detection; TMB (3, 3´, 5, 5´-tetramethylbenzidene)

Membrane Substrate and 4 CN (4-chloro-1-naphthol)

Membrane Substrate. TMB substrate offers the most

sensitive colorimetric detection for blotting applica-

tions and is recommended when high signal and low

background are required. 4 CN substrate is highly

reliable and is notable for its characteristically low

background.

• TMB Membrane Substrates (1- and (3-Component)

In the presence of peroxidase conjugate, KPL’s TMB

Membrane Substrates produce a stable, dark blue pre-

cipitate at the site of a positive reaction, enabling

detection of as little as 50 pg of peroxidase. Our TMB

substrates are available as 1- and 3- component solu-

tions. 1-component TMB Peroxidase Substrate is ready

to use and contains hydrogen peroxide and buffer.

Our 3-component TMB contains TMB in an organic

base, H2O2 in a citric acid buffer and a third compo-

nent, TMB Membrane Enhancer. It provides sensitivi-

ty equivalent to 1-component TMB at an economic

price. The substrate may be adapted as a soluble sub-

strate for ELISA by omitting TMB Membrane

Enhancer. Both substrates perform equally well on

nitrocellulose and PVDF membranes.

• 4 CN Peroxidase Substrate (2-Component)

4 CN (4-chloro-1-napthol) Peroxidase Membrane

Substrate produces a purple precipitate at the reaction

site in the presence of peroxidase conjugates. Due to

its low background, 4 CN presents a desirable alter-

native to other, more sensitive substrates when back-

ground poses a problem.

Sensitivity, reliability and affordability!

SEE MORE with KPL!

HRP Membrane Color Substrates

4 CN produces adistinct, blue-pur-ple precipitate andextremely lowbackground.

TMB produces ablue precipitateand providesgreater sensitivitythan 4 CN.

4 CN TMB

•

•

Benefits of KPL’s HRP Membrane Substrates


To order or for more information on KPL’s line of protein detection products, contact

us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com.


50-77-18 TMB Membrane Substrate (1-Component) 100 mL


50-77-04 TMB Membrane Substrate (1-Component) 1 L


50-77-01 TMB Membrane Enhancer 40 mL

50-73-00 4 CN Substrate System (2-Component) 600 mL

50-73-04 4 CN Substrate System (2-Component) 2700 mL

Protein Detector TMB Western Blot Kit

54-11-50 Protein Detector TMB Western Blot Kit 2500 cm2

Detector Block

71-83-00 Detector Block 240 mL

KPL provides a broad range ofassay support reagents forWestern blot detection.Conjugate stabilizers, diluents,block and wash solutions areoffered in convenient stable liq-uid form to ensure accuratereproducible results, blot afterblot. Our Detector Block isideal for use as a diluent/blocksolution for Western blots,Southern blots and Northernblots on a variety of mem-branes.

• DetectorTM Block

Detector Block is recommend-ed as a general blocking agentin membrane-based detectionassays. It is especially usefulwhen traditional blocking solu-tions (i.e., milk, BSA, casein)reduce sensitivity or do not ade-quately block background.Detector Block can be used asboth a blocking solution and aconjugate diluent in membraneapplications.

Support Reagents


Phone: 800.638.3167

Fax: 301.948.0169

www.kpl.com

Protein Detector is a trademark of KPL.©2008 KPL, Inc. All rights reserved.ML353-01

SEE MORE with KPL!


4 CN Peroxidase Substrate

TMB Peroxidase Membrane Substrate

• Most sensitive colorimetric substrate for blotting• Delivers results at the picogram level• Recommended for both kinetic and endpoint ELISA • Contains no harmful organic solvents; safer and more sensitive

than DAB or AEC.• 1-Component TMB-no preparation required.

Ensures more consistent results• 3-Component TMB-same performance as 1-Component TMB

at an economic price. • Blots remain stable with minimal fading

• Easy mix-and-use, two-component solutions• Very low background, moderate sensitivity• High contrast image ideal for photography and publication

Lighten your work load with KPL’s new line of quality

buffers. Offered as convenient liquid concentrates, they

eliminate the need for time-consuming buffer preparation.

KPL buffers are manufactured and carefully controlled for

quality and consistent performance with our ISO 9001:2008-

registered quality management system. All buffers are con-

ductivity controlled. KPL offers a variety of buffers designed

for use in general immunoassay applications such as Western

blotting, gel electrophoresis, ELISA and sample preparation.

Save preparation time with KPL’s new line of quality buffers!

Immunoassay Buffers

SEE MORE with KPL!

Description 1X Composition Applications

10X Tris-Glycine 25 mM Tris, 192 mM glycine. For preparing a standard Western blot transfer buffer Transfer Buffer pH 8.3 (Towbin) and as a gel electrophoresis buffer for native

Tris-glycine gels without SDS. .

10X Tris-Glycine-SDS 25 mM Tris, 192 mM glycine, Running buffer for sodium dodecyl sulfate – polyacrylamide0.1% SDS. pH 8.3 gel electrophoresis (SDS-PAGE) of proteins.

10X Phosphate-Buffered 10 mM Phosphate,150 mM NaCl. Wash buffer for general immunoassay applicationsSaline (PBS) pH 7.4 and sample preparation. Where applicable, Tween 20

is a nonionic detergent that reduces nonspecificPhosphate-Buffered 10 mM Phosphate, 150 mM NaCl, binding and protein-protein interaction during wash steps.Tween 20, (PBST) 0.05% Tween 20. pH 7.4

10X Tris-Buffered Saline 50 mM Tris, 150 mM NaCl. (TBS) pH 7.6

10% Tween 20 10% Tween 20 Nonionic detergent additive for PBS and Western blotting wash solutions. Reduces nonspecific binding and protein- protein interactions.

See reverse side.


51-10-01 10X Tris-Glycine Transfer Buffer 1 L

51-10-02 5 L

51-11-01 10X Tris-Glycine SDS Buffer 1 L

51-11-02 5 L

51-13-01 10X Phosphate-Buffered Saline (PBS) 1 L

51-13-02 5 L

51-14-01 10X Phosphate-Buffered Saline with 200 mL

51-14-02 Tween 20 (PBST) 1 L

51-17-01 10X Tris-Buffered Saline (TBS) 1 L

51-17-02 5 L

51-12-01 10% Tween 20 200 mL

51-12-02 1 L

51-20-01 5% SDS 200 mL

50-86-05 20X SSC 1 L

51-15-01 10X Dulbecco's PBS (D-PBS) 1 L

51-15-02 5 L

51-16-01 10X Dulbecco's PBS with Tween 20 (D-PBST) 200 mL

51-16-02 1 L

51-18-01 10X Tris Buffered Saline with 0.5% Tween 20 200 mL

51-18-02 (TBST) 1 L

51-19-01 10X Tris Buffered Saline with 10% Tween 20 200 mL

51-19-02 (TBST) 1 L


To order or for more information on KPL’s full line of protein detection products, contact us at800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com.

Other Immunoassay Support

Reagents Available from KPL

ELISA and Western Blotting

Applications:

• AP and HRP Conjugate Stabilizers

• 10% BSA Diluent/Blocking Solution

Concentrate

• Milk/Diluent Blocking Solution

Concentrate

• 20X Wash Solution Concentrate

Western Blotting Applications:

• Detector Block

• Biotin Wash Kit

ELISA Applications:

• Coating Solution Concentrate

• Stop Solutions

Immunoassay Buffers


Phone: 800.638.3167

Fax: 301.948.0169

www.kpl.com

ML363-03

©2009 KPL, Inc. All rights reserved.SEE MORE with KPL!


KPL Quality Buffers (cont’d)

Take the time and effort out of your buffer preparation. Choose from our line of popular buffers andenjoy the benefits - convenience, reliability and confidence - that KPL’s pretested, quality buffersbring to your research.

Description 1X Composition Applications

5% SDS 5% Sodium Detergent surfactant used in preparing proteins for Dodecyl Sulfate sodium dodecyl sulfate – polyacrylamide gel electro-

phoresis (SDS-PAGE) and as an additive to transfer buffers in Western blotting. SDS increases the elution rate of proteins from the gel.

20X SSC 15 mM sodium citrate, Used in a variety of molecular biology applications. 150 mM NaCl Facilitates transfer of nucleic acids to membranes. in DEPC water. pH 7.0

10X Dulbecco's 8.5 mM sodium phosphate, For sample preparation and as a wash buffer in general PBS (D-PBS) 1.5 mM potassium phosphate, immunoassay, tissue, and cell culture applications.

137 mM NaCl. pH 7.4 Not formulated with magnesium or calcium salts. Where applicable, Tween 20 is a non-ionic detergent that reducesnonspecific binding.

10X Dulbecco's 8.5 mM sodium phosphate,with Tween 20. 1.5 mM potassium phosphate,(D-PBST) 137 mM NaCl, 2.7 mM KCl.

0.05% Tween 20. pH 7.4

10X Tris-Buffered 50 mM Tris, 150 mM NaCl, As a general wash buffer in immunoassay applications. Saline with 0.5% 0.05% Tween 20. pH 7.6 Tween 20 is a non-ionic detergent that reduces nonspecific Tween 20 (TBST) binding. The 10% Tween 20 formulation provides a more

stringent wash than standard TBST formulations. 10X Tris-Buffered 50 mM Tris, 150 mM NaCl,Saline with 10% 1.0% Tween 20. pH 7.6 Tween 20 (TBST)

KPL’s new ReserveAPTM Alkaline Phosphatase

(AP)-labeled antibody conjugates exhibit high

potency and consistent performance in

immunoassays. These conjugates are the result

of advances in our conjugation technology and

offer higher signal than our current line of AP

conjugates while meeting the same standards

for low background, stability and reproducibili-

ty. They are intended for demanding

immunoassays that require high detection sen-

sitivity, including ELISA, Western blotting and

immunohistology.

Higher Potency

ReserveAP Conjugates are affinity purified and

conjugated to the highest grade of alkaline

phosphatase. In our studies, they generate two-

to-three fold higher values than our current

line of AP conjugates in ELISA and outperform

AP conjugates offered by other manufacturers.

Higher conjugate dilutability is also observed

without loss of linearity, enabling precious

antigen or primary antibody conservation.

Consistent Performance

Reproducible antibody conjugation and consis-

tent performance are verified according to our

ISO 9001:2000-registered quality assurance

system. Lot consistency studies in which three

lots were studied by ELISA indicated minimal

variability.

Excellent Value

ReserveAP Conjugates provide high perform-

ance at an economical price.

Spice up your assay with our red hot high potency ReserveAPTM Conjugates!

ReserveAPTM Conjugates

SEE MORE with KPL!

ReserveAPTM Conjugates Ordering Information


0751-1001 Goat Anti-Human IgA (α) 1.0 mg

0751-1002 Goat Anti-Human IgG (γ) 1.0 mg

0751-1003 Goat Anti-Human IgM (µ) 1.0 mg

0751-1004 Goat Anti-Human IgE (ε) 1.0 mg

0751-1006 Goat Anti-Human IgG (H+L) 1.0 mg

0751-1007 Goat Anti-Human IgA+IgG+IgM (H+L) 1.0 mg

2151-1002 F(ab’)2 Anti-Human IgG (γ) 0.5 mg

4751-1002 Goat Anti-Human IgG (γ) liquid 1.0 mg

4751-1003 Goat Anti-Human IgM (µ) liquid 1.0 mg

4751-1006 Goat Anti-Human IgG (H+L) liquid 1.0 mg

0751-1802 Goat Anti-Mouse IgG (γ) HSA 1.0 mg

0751-1803 Goat Anti-Mouse IgM (µ) HSA 1.0 mg

0751-1806 Goat Anti-Mouse IgG (H+L) HSA 1.0 mg

0751-1809 Goat Anti-Mouse IgG +IgM (H+L) HSA 1.0 mg

4751-1802 Goat Anti-Mouse IgG (γ) HSA, liquid 1.0 mg

4751-1806 Goat Anti-Mouse IgG (H+L) HSA, liquid 1.0 mg

151-18-01 Goat Anti-Mouse IgA (α) HSA 0.5 mg

0751-1807 Goat Anti-Mouse IgA+IgG+IgM (H+L) HSA 1.0 mg

0751-1506 Goat Anti-Rabbit IgG (H+L) 1.0 mg

0751-1516 Goat Anti-Rabbit IgG (H+L) HSA 1.0 mg

4751-1506 Goat Anti-Rabbit IgG (H+L), liquid 1.0 mg

4751-1516 Goat Anti-Rabbit IgG (H+L) HSA, liquid 1.0 mg

HSA=Human Serum Adsorbed

AP Substrates


Phone: 800.638.3167

Fax: 301.948.0169

www.kpl.com


ML349-04

For research use only.©2007 KPL, Inc. All rights reserved.

SEE MORE with KPL!

Visit our website at www.kpl.com for a complete listing of ReserveAP conjugates. To

order or for more information on KPL’s protein research products, contact us at

800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

KPL offers a range of sensitive sub-

strates for the detection and quantifi-

cation of phosphatase (AP) activity.

They provide a choice of intense col-

ors for ELISA and blotting applica-

tions.

ELISA

• FirePhosTM AP Microwell

Substrate

• BluePhos® AP Microwell

Substrate

• pNPP Phosphatase Substrate

Blotting

• FirePhos AP Membrane

Substrate

• BCIP/NBT Phosphatase Substrate

• PhosphaGLO AP Substrate

• PhosphaGLOTM Reserve AP

Substrate

Whichever substrate you choose,

enjoy the benefits of excellent signal-

to-noise with higher sensitivity and

lower background than that of other

AP substrates. Visit www.kpl.com

for more information.

Chemiluminescent ELISA: A con-stant amount of antigen (mouse IgG)was coated onto a microtiter plate.The plate was probed with varyingconcentrations of goat anti-mouseIgG (H+L) AP conjugate, includingKPL’s standard AP conjugate andReserveAP™ conjugate, followed byKPL’s PhosphaGLO™ Chemilum-inescent AP Substrate.

Conclusion: The results indicate that ReserveAP™ conjugate offers superior performance to our stan-dard AP conjugate with a ~3-fold lower amount of conjugate required for detection.

KPL’s DyLightTM conjugates offer a brilliant

choice in a variety of multicolor detection ap-

plications, including fluorescence microscopy,

flow cytometry, Western blotting, ELISA and

array platforms. Our affinity purified antibod-

ies combined with a series of outstanding

DyLight dyes provide superior performance

over conventional CyDyeTM fluors, fluorescein

and rhodamine, with performance comparable

to that of Alexa Fluor® dyes (Figure 1). Enjoy

these advantages when you switch to KPL’s

DyLight Conjugates:

KPL offers eight DyLight dyes, including 405,

488, 549, 594, 633, 649, 680 and 800 with

well-differentiated excitation and emission

spectra. Our extensive line of over 170 DyLight

conjugates is available across a range of animal

species immunoglobulin, including human,

mouse, rabbit, rat, other species and strepta-

vidin. See back cover to find out what sets KPL’s

antibodies apart.

SEE MORE with KPL!

KPL’s DyLightTM Conjugates - A Brilliant Choice!

DyLightTM Fluorescent Conjugates

Image above: HMVEC-L primary endothelial cells. F-actin detected with DY554-Phalloidin (rendered green). Microtubules detected with anti-tubulinantibody and DyLight 649 conjugated goat anti-mouse IgG (red). Nuclei detected with DAPI (blue)

Figure 1. KPL DyLight 649 conjugates demonstrate comparablefluorescense intensity and photostability to Alexa 647 conjugatesin a FLISA.

•• High sensitivity, bright signal enable conjugate

conservation

•• More photostable than Cy, FITC; comparable to Alexa

•• High signal-to-noise ratios; low background

•• Wide selection of specific antibodies

•• Consistent, lot-to-lot performance

•• Narrow emission spectra enable specific, multicolor analysis

•• Compatible with a variety of buffers.

Antigen Concentration (ng/mL)

Comparison of DyLight 649 and Alexa 647Conjugates in a Performance Assay

Emission DyLight Dye Ex/Em(nm) Replaces

DyLight 405 400/420 Alexa 405 and Cascade Blue

DyLight 488 493/518 Alexa 488, Fluorescein and FITC

DyLight 549 550/568 Alexa 546, Alexa 555, Cy3, TRITC

DyLight 594 593/618 Alexa 594, Texas Red

DyLight 633 638/658 Alexa 633

DyLight 649 646/674 Alexa 647, Cy5

DyLight 680 682/715 Alexa 680, Cy5.5

DyLight 800 770/794 IRDye 800

KPL DyLight conjugates provide a choice of outstanding flu-

orescent conjugates with absorption spectra ranging from

405 nm to 800 nm. Their emission profiles correspond to

those of other commonly used fluorophores such as Alexa,

FITC and the Cy dyes. Narrow, defined emission spectra en-

able the use of multiple fluorescence labeling and simultane-

ous identification of several target molecules in the same

sample. Light output is comparable to IRDye and Alexa and

more intense than Cy Dyes, FITC or TRITC. DyLight dyes

demonstrate resistance to photobleaching, resulting in excel-

lent photostability as well as high solubility in aqueous solu-

tions across a range of pH values.

Anti-Mouse IgG (γ), HSA 072-03-18-02 072-04-18-02 072-05-18-02

Anti-Mouse IgG (H+L), HSA 072-08-18-06 072-03-18-06 072-04-18-06 072-09-18-06 072-10-18-06 072-05-18-06 072-06-18-06 072-07-18-06

Anti-Mouse IgG (H+L), HSA, 0.1 mg 042-08-18-06 042-03-18-06 042-04-18-06 042-09-18-06 042-10-18-06 042-05-18-06 042-06-18-06 042-07-18-06

Anti-Mouse IgG (H+L), RbSA, HSA 072-03-18-18 072-04-18-18 072-05-18-18 072-06-18-18 072-07-18-18

Anti-Mouse IgM (µ), HSA 072-03-18-03 072-04-18-03 072-05-18-03

Anti-Mouse IgG+IgM (H+L), HSA 072-03-18-09 072-04-18-09 072-05-18-09

F(ab’)2 Anti-Mouse IgG (γ), HSA 202-03-18-02 202-04-18-02 202-05-18-02

F(ab’)2 Anti-Mouse IgG (H+L), HSA 202-08-18-06 202-03-18-06 202-04-18-06 202-09-18-06 202-10-18-06 202-05-18-06 202-06-18-06 202-07-18-06

Anti-Human IgG (H+L) 072-08-10-06 072-03-10-06 072-04-10-06 072-09-10-06 072-10-10-06 072-05-10-06 072-06-10-06 072-07-10-06

Anti-Human IgG (H+L), 0.1 mg 042-08-10-06 042-03-10-06 042-04-10-06 042-09-10-06 042-10-10-06 042-05-10-06 042-06-10-06 042-07-10-06

Anti-Human IgG (γ) 072-03-10-02 072-04-10-02 072-05-10-02

Anti-Human IgM (µ) 072-03-10-03 072-04-10-03 072-05-10-03

F(ab’)2 Anti-Human IgG (γ) 202-03-10-02 202-04-10-02 202-05-10-02

F(ab’)2 Anti-Human IgM (µ) 202-03-10-03 202-04-10-03 202-05-10-03

F(ab’)2 Anti-Human IgG (H+L) 202-08-10-06 202-03-10-06 202-04-10-06 202-09-10-06 202-10-10-06 202-05-10-06 202-06-10-06 202-07-10-06

Anti-Rabbit IgG (H+L) 072-08-15-06 072-03-15-06 072-04-15-06 072-09-15-06 072-10-15-06 072-05-15-06 072-06-15-06 072-07-15-06

Anti-Rabbit IgG (H+L), 0.1 mg 042-08-15-06 042-03-15-06 042-04-15-06 042-09-15-06 042-10-15-06 042-05-15-06 042-06-15-06 042-07-15-06

Anti-Rabbit IgG (H+L), HSA 072-03-15-16 072-04-15-16 072-05-15-16 072-06-15-16 072-07-15-16

F(ab’)2 Anti-Rabbit IgG (H+L), HSA 202-08-15-16 202-03-15-16 202-04-15-16 202-09-15-16 202-10-15-16 202-05-15-16 202-06-15-16 202-07-15-16

Anti-Rat IgG (H+L) 072-08-16-06 072-03-16-06 072-04-16-06 072-09-16-06 072-10-16-06 072-05-16-06 072-06-16-06 072-07-16-06

Anti-Rat IgG (H+L), 0.1 mg 042-03-16-06

Anti-Guinea Pig IgG (H+L) 072-03-17-06 072-04-17-06 072-05-17-06 072-06-17-06 072-07-17-06

Anti-Chicken IgG (H+L) 072-08-24-06 072-03-24-06 072-04-24-06 072-09-24-06 072-10-24-06 072-05-24-06 072-06-24-06 072-07-24-06

Anti-Dog IgG (H+L) 072-03-19-06 072-04-19-06 072-05-19-06 072-06-19-06 072-07-19-06

Anti-Goat IgG (H+L) 072-08-13-06 072-03-13-06 072-04-13-06 072-09-13-06 072-10-13-06 072-05-13-06 072-06-13-06 072-07-13-06

Anti-Horse IgG (H+L) 072-03-21-06 072-04-21-06 072-05-21-06 072-06-21-06 072-07-21-06

Rabbit Anti-Sheep IgG (H+L) 072-03-23-06 072-04-23-06 072-05-23-06 072-06-23-06 072-07-23-06

Anti-Swine IgG (H+L) 072-03-14-06 072-04-14-06 072-05-14-06 072-06-14-06 072-07-14-06

Streptavidin 072-08-30-00 072-03-30-00 072-04-30-00 072-09-30-00 072-10-30-00 072-05-30-00 072-06-30-00 072-07-30-00

Streptavidin, 0.1 mg 042-08-30-00 042-03-30-00 042-04-30-00 042-09-30-00 042-10-30-00 042-05-30-00 042-06-30-00 042-07-30-00

DyLightTM FluorescentConjugates

DyLight Conjugated Affinity PurifiedAntibodies and Streptavidin

nm = nanometersIR = infrared

Description DyLight 405* DyLight 488 DyLight 549 DyLight 594* DyLight 633* DyLight 649 DyLight 680 DyLight 800

SEE MORE with KPL!

*Coming soon!

Blue

Green

Yellow

Orange

Red

Red

Near IR

Infrared


HSA = human serum adsorbedRbSA = rabbit serum adsorbedDyLight antibody conjugates are made in goat exceptanti-goat and anti-sheep antibodies are made in rabbit.Supplied in 1.0 mg lyophilized form except select 0.1mg sizes.

DyLight Dyes Emission Spectra

Near Infrared and Infrared Fluorophores DyLight680 and 800 Conjugates Offer Sensitive MulticolorImaging in Western Blotting

DyLight 680 and 800 dyes emit in the near infrared and

infrared ranges of the light spectrum respectively and are

ideal for multicolor protein detection in Western blot-

ting. Unlike fluorescent conjugates that emit in the visi-

ble range, DyLight 680 and 800 conjugates provide a

unique set of advantages:

•• Secondary antibodies with defined specificity and

sensitivity

•• Brighter signal than visible fluorescence

•• Virtually no background autofluorescence from

membranes or most biological specimens

Effective Alternative to Chemiluminescence

•• Broader dynamic range than chemiluminescence

•• Quantitation accuracy superior to traditional

methods

•• Easy to assay multiple proteins simultaneously on

one Western blot

•• Cost-effective - eliminates need for chemilumin-

escence substrates, film and darkroom

•• Clean results - no bleeding from consecutive lanes

Figure 2. DyLight 680-anti mouse IgG and DyLight800 anti-rabbit IgG secondary antibody conjugatesprovide low background and high signal in two-colorWestern blot detection of tubulin and TNFα. Mem-brane was imaged with the LI-COR Odyssey InfraredImaging System.

Near Infrared Fluorescent Imaging with SecondaryAntibodies labeled with DyLight 680 and 800

SEE MORE with KPL! 1.800.638.3167 www.kpl.com

Gaithersburg, MD

Phone: 800.638.3167/301.948.7755

Fax: 301.948.0169

www.kpl.com

SEE MORE with KPL!


DyLightTM FluorescentConjugates

KPL Antibodies-What Sets Them Apart

ML356-02For research use only.DyLight is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries. Cy and Cy Dye are trademarks of GE Healthcare.Alexa Fluor is a registered trademark of Invitrogen.2009 KPL, Inc. All rights reserved.

Rat hippocampal neuronsWCS stained with; DyLight 488(green). MAP2 stained withDyLight 549 (red).

Mouse primary cortical neuronsMCx WCS stained with DyLight 549(red). Synaptophysin stained withDyLight 488 (green).

HMVEC-L primary endothelial cellsF-actin detected with DY554-Phal-loidin (rendered green). Micro-tubules stained with DyLight 649(red). Nuclei detected with DAPI(blue).

NIH 3T3 cells F-actin detected with DY554-Phal-loidin (rendered green). Micro-tubules stained with DyLight 649(red). Nuclei detected with DAPI(blue)

A549 cells Cytokeratin stained with DyLight680 (pseudocolored white).Lamin A stained with DyLight549 (pseudocolored red).

Immunofluorescence using DyLight Conjugates

See bright fluorescence and low background with KPL DyLight conjugates in im-

munohistology applications. The winning combination of DyLight dyes and KPL

purified antibodies enables multicolor labeling of two or more targets with similar

intensity and photostability to Alexa dyes without the limitations of fluorescein

and CyDyes.

To order or for more information on KPL’s line of unlabeled and conjugated

affinity purified antibodies, contact us at 800.638.3167/301.948.7755,

fax 301.948.0169 or visit www.kpl.com.

In 1979, KPL pioneered the productionof large-scale affinity purification andwas the first company to commercializeaffinity purified secondary antibodies.Rigorous standards throughout the anti-body production process make ourantibodies standout in the marketplace.Many manufacturers cut corners by be-ginning with inferior serum and extract-ing the useful antibody towards the endof their process. KPL spends considerableeffort developing and purifying its ownimmunogen formulation to generate theantiserum, because pure immunogenresults in a more potent and specificantibody prior to any purification steps.

Further, ISO 9001:2008-certified qualityprocedures are carried out at more thansix different stages of the antibody pro-duction cycle, and material that does notmeet our high standards for potency andcross-reactivity is rejected. Our latestage purification process has been con-tinually refined since 1979 and relies ona one of a kind custom column resinwhich is uniquely suited to our antibodymanufacturing process.

Finally, our process of pooling antiserumfrom multiple animals tempers naturalserum variability, minimizing variancesfrom animal-to-animal. The result ismore standardized large-scale antibodylots with increased consistency. KPL’s ex-perience, innovative processes, and at-tention to detail result in high-performance antibodies that are unique.

Cy Dyes offer intense fluorescence when coupled

with KPL’s affinity purified antibodies and strepta-

vidin. The sensitivity and reproducibility of KPL

antibodies combined with the brightness of Cy3 and

Cy5 dyes produce an exceptional set of conjugates

ideal for multiple labeling experiments. Dye/protein

ratios have been established to ensure optimal fluo-

rescence with minimal background. They present

maximum excitation/emission spectra at 550/570

nm (Cy3) and 650/670 nm (Cy5).

Cy dyes are excellent alternatives to most other fluor-

rescein dyes as they are brighter and offer greater

photostability. Cy Dye conjugates are used in both

visual and image analysis fluorescent microscopy

and in situ hybridization. Cy3 conjugates are ideally

used in visual color applications, whereas Cy5 con-

jugates emit in the far red spectrum and are not easi-

ly visualized.

Benefits of KPL Cy Dye Conjugates

High performance conjugates – optimized dye-protein ratios ensure high signal-to-noise.

Intense fluorescence – offers greater sensitivity than TRITC conjugates.

Narrow emission spectra – enables sensitive, multi-color analysis.

Excellent photostability – more photostable than TRITC conjugates.

Consistent performance – minimal lot-to-lot vari-ation reduces need for assay optimization.

Buffer stability – after rehydration, conjugates are stable at pH 4-9.

Instrument compatibility – excitation and emis-sion spectra correspond with standard filter sets and laser settings.

KPL CyTM Dye Conjugates- Improve your assay and brighten your day!

SEE MORE with KPL!

Cy3 and Cy5 Conjugates

Fluo

resc

ence

Wavelength (nm)

Cy3 Cy5ExEm

Image above: Confocal fluorescence micrograph of HeLa cells stained with monoclonal antibody against mitochondria enzyme and Cy3-

conjugated anti-mouse antibody (red); rabbit polyclonal antibody to histones in DNA and Cy5-conjugated anti-rabbit antibody (blue).


To order or for more information on KPL’s line of unlabeled and conjugated

affinity purified antibodies, contact us at 800.638.3167 / 301.948.7755, fax

301.948.0169 or visit us at www.kpl.com.

Cy3 and Cy5 Conjugates

ML368-01For research use only.Cy Dye is a trademark of GE Healthcare.©2009 KPL, Inc. All rights reserved.

Cytomegalovirus-infected cellsdetected with a biotinylated CMVprobe and the DNADetectorTM

Fluorescent in situ Hybridization Kitusing Cy3-Strept-avidin and DAPI.

Signal DetectionCy3 is excited maximally at 550 nmand fluoresces maximally at 570 nm.It is excited to about 50% of maxi-mum with an argon laser (514 nm or528 nm lines), or to about 75% ofmaximum with a helium/neon laser(543 nm line) or mercury lamp (546nm line).

Cy5 is excited maximally at 650 nmand fluoresces maximally at 670 nm.It is excited to about 98% of maxi-mum with a krypton/argon laser (647nm line) or to about 63% of maxi-mum with a helium/neon laser (633nm line). Cy5 produces minimal auto-fluorescence of biological specimensin this region of the spectrum.

A confocal microscope equipped withthe appropriate laser for excitationand a far-red detector enable doublelabeling with Cy3 and Cy5.


Phone: 800.638.3167

Fax: 301.948.0169

www.kpl.com


SEE MORE with KPL!

Anti-Mouse IgG (γ), HSA 072-01-18-02 072-02-18-02

F(ab’)2 Anti-Mouse IgG (γ), HSA 202-01-18-02 202-02-18-02

Anti-Mouse IgG (H+L), HSA 072-01-18-06 072-02-18-06

F(ab’)2 Anti-Mouse IgG (H+L), HSA 202-01-18-06 202-02-18-06

Anti-Mouse IgG (H+L), RbSA, HSA 072-01-18-18 072-02-18-18

Anti-Mouse IgM (µ), HSA 072-01-18-03 072-02-18-03

Anti-Mouse IgG+IgM (H+L), HSA 072-01-18-09 072-02-18-09

Anti-Rabbit IgG (H+L) 072-01-15-06 072-02-15-06

F(ab’)2 Anti-Rabbit IgG (H+L), HSA 202-01-15-16 202-02-15-16

Anti-Rabbit IgG (H+L), HSA 072-01-15-16 072-02-15-16

Anti-Rat IgG (H+L) 072-01-16-06 072-02-16-06

F(ab’)2 Anti-Human IgG (H+L) 202-01-10-06 202-02-10-06

Anti-Human IgG (γ) 072-01-10-02 072-02-10-02

F(ab’)2 Anti-Human IgG (γ) 202-01-10-02 202-02-10-02

Anti-Human IgM (µ) 072-01-10-03 072-02-10-03

F(ab’)2 Anti-Human IgM (µ) 202-01-10-03 202-02-10-03

Anti-Guinea Pig IgG (H+L) 072-01-17-06 072-02-17-06

Anti-Chicken IgG (H+L) 072-01-24-06 072-02-24-06

Anti-Horse IgG (H+L) 072-01-21-06 072-02-21-06

Anti-Swine IgG (H+L) 072-01-14-06 072-02-14-06

Anti-Dog IgG (H+L) 072-01-19-06 072-02-19-06

Anti-Sheep IgG (H+L) 072-01-23-06 072-02-23-06

Anti-Goat IgG (H+L) 072-01-13-06 072-02-13-06

Streptavidin 072-01-30-00 072-02-30-00

Description Cy3 Cy5

HSA=human serum adsorbed RbSA=rabbit serum adsorbedCy Dye antibody conjugates are made in goat except anti-goat and anti-sheep antibodiesmade in rabbit. Supplied in 1 mg lyophilized form.

Microplates were coated with serially diluted mouse IgG at the indicated concentrations. Conjugates at aconcentration of 0.01 mg/mL were applied and incubated for 30 minutes. Fluorescence was measured witha Perkin Elmer VICTOR 3 Multilabel Plate Reader.

Excellent PerformanceAs demonstrated below KPL Cy3- and Cy5-labeled conjugates produce brighter fluorescence thanthose of other suppliers.

Since 1979 KPL has provided quality affinity

purified antibodies to researchers worldwide.

Over the years we have refined our production

process to provide antibodies with high potency

and consistent performance in immunoassays.

From the start KPL gives careful consideration

to immunogen preparation, using a highly

purifed formulation to generate antiserum. KPL

pools antiserum from multiple animals to

reduce natural animal to animal serum variabili-

ty. During the purification process our ISO

9001:2008-certified quality procedures impose

rigorous standards for potency and cross-reac-

tivity. The result is standardized antibody lots

with excellent reproducibility.

Our extensive line of peroxidase (HRP) conju-

gates is available across a range of animal

species, including human, mouse, rabbit and rat

antibodies, as well as other animal species and

streptavidin. They are affinity purified and in

some cases further adsorbed to minimize cross-

reactivity between animal species or shared

reactivity with other immunoglobulin classes.

HRP-labeled F(ab’)2 fragment antibodies are

offered for assays requiring extremely low back-

ground and absence of F(c)-mediated binding.

KPL reacts HRP of the highest quality with

affinity purified antibodies and streptavidin

using the periodate method of Nakane and

Kawaoi. Special features of HRP include:

• faster catalytic rate than alkaline phosphatase

• generates more product in shorter incubation

times

• provides maximum sensitivity, low nonspe-

cific binding

• ideal for ELISA, Western blotting and

immunohistology applications.

KPL Peroxidase Conjugates: Time-tested, Sensitive and Reliable

Peroxidase Conjugates

SEE MORE with KPL!

Peroxidase Conjugates Ordering Information(Partial listing)

Peroxidase Conjugates


Phone: 800.638.3167

Fax: 301.948.0169

www.kpl.com


For research use only.©2009 KPL, Inc. All rights reserved.ABTS is a registered trademark of Roche Biochemicals.ML371-01

SEE MORE with KPL!

Visit our website at www.kpl.com for a complete listing of HRP-labeled antibodies.

To order or for more information on KPL’s protein research products, contact us at

800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

KPL offers a range of sensitive

substrates for use with HRP conju-

gates. They provide a choice of

intense colors for ELISA, blotting

and cell staining applications.

ELISA

• ABTS® 1- and 2-Component

Microwell Peroxidase Substrates

• SureBlueTM TMB Peroxidase

Substrate

• SureBlue ReserveTM TMB

Peroxidase Substrate

• TMB Peroxidase Substrate

Blotting

• 4 CN Peroxidase Substrate

• TMB Membrane Peroxidase

Substrate

• LumiGLO®Chemiluminescent

Substrate

• LumiGLO ReserveTM

Chemiluminescent Substrate

Whichever substrate you choose,

enjoy the benefits of excellent signal-

to-noise and reproducibility.


04-10-06 HRP-labeled Goat Anti-Human IgG (H+L) 0.1 mg

04-10-17 HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L), MSA 0.1 mg

04-10-20 HRP-labeled Goat Anti-Human IgG (Fc) 0.1 mg

074-1002 HRP-labeled Goat Anti-Human IgG (γ) 1.0 mg

074-1003 HRP-labeled Goat Anti-Human IgM (μ) 1.0 mg

074-1004 HRP-labeled Goat Anti-Human IgE (ε) 1.0 mg

074-1006 HRP-labeled Goat Anti-Human IgG (H+L) 1.0 mg

074-1007 HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L) 1.0 mg

14-10-01 HRP-labeled Goat Anti-Human IgA (α) 0.5 mg

214-1002 HRP-labeled F(ab’)2 Goat Anti-Human IgG (γ) 0.5 mg

214-1003 HRP-labeled F(ab’)2 Goat Anti-Human IgM (μ) 0.5 mg

214-1006 HRP-labeled F(ab’)2 Goat Anti-Human IgG (H+L) 0.5 mg

474-1002 HRP-labeled Goat Anti-Human IgG (γ), Liquid 1.0 mL

474-1003 HRP-labeled Goat Anti-Human IgM (μ), Liquid 1.0 mL

474-1006 HRP-labeled Goat Anti-Human IgG (H+L), Liquid 1.0 mL

04-18-06 HRP-labeled Goat Anti-Mouse IgG (H+L), HSA 0.1 mg

04-18-15 HRP-labeled Goat Anti-Mouse IgG (H+L), RtSA, HSA 0.1 mg

04-18-18 HRP-labeled Goat Anti-Mouse IgG (H+L), RbSA, HSA 0.1 mg

074-1802 HRP-labeled Goat Anti-Mouse IgG (γ), HSA 1.0 mg

074-1803 HRP-labeled Goat Anti-Mouse IgM (μ), HSA 1.0 mg

074-1806 HRP-labeled Goat Anti-Mouse IgG (H+L), HSA 1.0 mg

074-18-061 HRP-labeled Goat Anti-Mouse IgG (H+L), XSA 1.0 mg

074-1807 HRP-labeled Goat Anti-Mouse IgA+IgG+IgM (H+L), HSA 1.0 mg

074-1809 HRP-labeled Goat Anti-Mouse IgG+IgM (H+L), HSA 1.0 mg

14-18-01 HRP-labeled Goat Anti-Mouse IgA (α), HSA 0.5 mg

214-1802 HRP-labeled F(ab’)2 Goat Anti-Mouse IgG (γ), HSA 0.5 mg

214-1806 HRP-labeled F(ab’)2 Goat Anti-Mouse IgG (H+L), HSA 0.5 mg

474-1802 HRP-labeled Goat Anti-Mouse IgG (γ), HSA, Liquid 1.0 mL

474-1806 HRP-labeled Goat Anti-Mouse IgG (H+L), HSA, Liquid 1.0 mL

074-1506 HRP-labeled Goat Anti-Rabbit IgG (H+L) 1.0 mg

074-15-061 HRP-labeled Goat Anti-Rabbit IgG (H+L), XSA 1.0 mg

074-1516 HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA 1.0 mg

214-1516 HRP-labeled F(ab’)2 Goat Anti-Rabbit IgG (H+L), HSA 0.5 mg

474-1506 HRP-labeled Goat Anti-Rabbit IgG (H+L), Liquid 1.0 mL

474-1516 HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA, Liquid 1.0 mL

14-30-00 HRP-labeled Streptavidin 0.5 mg

474-3000 HRP-labeled Streptavidin, Liquid, Molecular Grade 1.0 mL


Catalog # Description Size

Protein Detector™ Western Blotting KitsEach kit includes anti-mouse and anti-rabbit conjugates, DetectorBlock, Wash Solution Concentrate and Substrate.

Phosphatase Chromogenic

55-11-50 BCIP/NBT Western Blot Kit 2500 cm2

Peroxidase Chromogenic

54-11-50 TMB Western Blot Kit 2500 cm2

Peroxidase Chemiluminescent54-12-50 LumiGLO® Western Blot Kit 2500 cm2

54-13-50 LumiGLO ReserveTM Western 2400 cm2

Blot Kit

Related Reagents and Kits

Antibody ConjugatesAll antibodies listed below are produced in goat.For a complete antibody listing, refer to KPL’s Product Catalog.

Phosphatase-labeled475-1006 Anti-Human IgG (H+L) 1.0 mL, liquid

475-1806 Anti-Mouse IgG (H+L), HSA 1.0 mL, liquid

475-1506 Anti-Rabbit IgG (H+L) 1.0 mL, liquid

Peroxidase-labeled474-1006 Anti-Human IgG (H+L) 1.0 mL, liquid

474-1806 Anti-Mouse IgG (H+L) HSA 1.0 mL, liquid

474-1506 Anti-Rabbit IgG (H+L) 1.0 mL, liquid

Biotin-labeled16-10-06 Anti-Human IgG (H+L) 0.5 mg

176-1006 Anti-Human IgG (H+L) 2.0 mg

Labeled Streptavidin474-3000 HRP-labeled 1.0 mL, liquid

475-3000 AP-labeled 1.0 mL, liquid

Catalog # Description Size

Substrates for Western Blotting

Phosphatase Colorimetric Substrates50-81-18 BCIP/NBT Substrate 100 mL

50-81-07 BCIP/NBT Substrate 600 mL

50-81-30 FirePhosTM Membrane AP Substrate 100 mL

50-81-40 FirePhos Membrane AP Substrate 400 mL

50-81-34 FirePhos Membrane AP Substrate 1000 mL

Phosphatase Chemiluminescent Substrates55-60-03 PhosphaGLOTM AP Substrate 30 mL

55-60-04 PhosphaGLO AP Substrate 100 mL

55-60-01 PhosphaGLO Reserve AP Substrate 30 mL

55-60-02 PhosphaGLO Reserve AP Substrate 100 mL

Peroxidase Chromogenic Substrates50-77-18 TMB Membrane Substrate 100 mL

50-77-03 TMB Membrane Substrate 200 mL

50-73-00 4 CN Substrate 600 mL

50-73-04 4 CN Substrate 2700 mL

Peroxidase Chemiluminescent Substrates54-61-02 LumiGLO Chemiluminescent Substrate 60 mL

54-61-00 LumiGLO Chemiluminescent Substrate 240 mL

54-61-01 LumiGLO Chemiluminescent Substrate 720 mL

54-71-00 LumiGLO Reserve Substrate Kit 2400 cm2

54-71-01 LumiGLO Reserve Substrate Kit 600 cm2

Assay Support Reagents50-84-00 Coating Solution Concentrate 50 mL

54-15-01 HRPStabilizer 200 mL

55-15-00 APStabilizer 200 mL

50-61-00 10% BSA Diluent/Blocking Solution 200 mLConcentrate

50-82-01 Milk Diluent/Blocking Solution 200 mLConcentrate

71-83-00 DetectorTM Block (5X) 240 mL

50-63-00 Wash Solution Concentrate (20X) 800 mL

50-63-06 Biotin Wash Solution Concentrate (10X) 200 mL

60-00-50 Biodyne® B Nylon Membrane 1 roll

HSA=human serum adsorbed.

To order or for more information,call KPL at 800.638.3167, 301.948.7755,Fax: 301.948.0169. www.kpl.comor contact your local sales partner.

ML282-04SEE MORE with KPL!

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