expressions - new england biolabs (uk) ltdavailable with and without competent e. coli, this...

NEB UKExpressions MAY

201

5

DNA Assembly and Cloning:NEBuilder HiFi DNA AssemblyGolden Gate Assembly

New Rapid PCR Cleanup Enzyme Set

CELL SIGNALING TECHNOLOGY®

CST™ SimpleChIP® Chromatin IP Kits and ChIP-validated Antibodies

NEBNext FFPE DNA Repair Mix

NEBNext Q5 Hot Start HiFi PCR Master Mix for improved amplification of NGS libraries

CONTENTS03 Rapid PCR Cleanup Enzyme Set – add NEB’s reagents directly to your PCR product for 100% sample recovery in 15 minutes.

04 The Next Generation of DNA Assembly: NEBuilder HiFi DNA Assembly – simple and fast Seamless Cloning of up to 12 fragments.

06 Introducing NEB Golden Gate Assembly Mix – Seamless Cloning that is particulary good with multiple fragments of high GC content and areas of repeats.

08 Improved amplification of NGS libraries – high yields, excellent GC coverage and ultra-high fidelity with NEBNext Q5 Hot Start HiFi PCR Master Mix.

09 Construct high-quality NGS libraries from FFPE DNA samples – NEBNext FFPE DNA Repair Mix.

10 Optimized Reagents Matter – SimpleChIP® Plus Enzymatic Chromatin IP Kits and ChIP-validated Antibodies from CST™.

100% 100% recycled paper, vegetable based inks.

Read Me, Recycle Me New England Biolabs (UK) Ltd75-77 Knowl Piece, Hitchin, Herts SG4 0TYTel: 0800 318486 | Email: [email protected]

FreeWith NEB

Free Delivery!

CELL SIGNALING TECHNOLOGY®, SIMPLECHIP®, XP® and E1L3N are registered trademarks of Cell Signaling Technology, Inc. CST™ is a trademark of Cell Signaling Technology, Inc. BIOANALYZER® is a registered trademark of Agilent Technologies, Inc. HYPERLADDER™ is a trademark of Bioline. TRACKIT™ is a trademark of Invitrogen™. INVITROGEN™ and TRACKIT™ are trademarks of Life Technologies, Inc. GENEART® is a registered trademark of Life Technologies, Inc. GIBSON ASSEMBLY® is a registered trademark of Synthetic Genomics, Inc. AMPURE® is a registered trademark of Beckman Coulter, Inc. ILLUMINA®, HISEQ® and MISEQ® are registered trademarks of Illumina, Inc.

Free Next Day Delivery for All UK Mainland Orders

new products

Quick-Load Purple DNA Ladders

• Sharp, Uniform Bands

• Easily Identifiable Reference Bands

• No UV Shadow

Be bright, go purple.

A. Lane 1. Invitrogen™ TrackIt™ 1 Kb Plus DNA Ladder; Lane 2. NEB Quick-Load Purple 2-log DNA Ladder; 1 µg per gel lane

B. The new Gel Loading Dye, Purple (6X) (Lane 1) included in the Quick-Load Purple 1 kb DNA Ladder does not cast a UV shadow over the underlying bands, unlike the Gel Loading Dye, Blue (6X) (Lane 2).

Ordering InformationPRODUCT NEB # SIZE PRICEQuick-load Purple 2-Log DNA Ladder N0550S 125-250 gel lanes £57

Quick-load Purple 100 kb DNA Ladder N0551S 125 gel lanes £64

Quick-load Purple 1kb DNA Ladder N0552S 125 gel lanes £46

30

90

10

100

0

80

Cost/

Lane

(Pen

ce)*

NEBQuick-Load

Purple 2-Log

BiolineHyperLadder™

1kb

* Small pack prices as of 02/15

InvitrogenTrackIt™ 1Kb Plus

20

Take Advantage of the Low Cost/Lane with NEB’s Quick-Load Purple 2-Log DNA Ladder

A. 1 2 B.

UV shadow

1 2

03

The NEB Rapid PCR Cleanup Enzyme Set consists of two recombinant enzymes, Exonuclease I (Exo I, #M0293) and Shrimp Alkaline Phosphatase (rSAP, #M0371). It allows for rapid and complete enzymatic degradation of residual PCR primers and dephosphorylation of dNTPs subsequent to PCR. The Rapid PCR Cleanup Enzyme Set enables direct downstream applications including sequencing, genotyping, cloning or SNP analysis without the need for additional purification steps. The Rapid PCR Cleanup Enzyme Set is added directly to the PCR product after thermal cycling and is 100% compatible with commonly used PCR reaction buffers.

Advantages:• Fast 15 minute protocol• Add directly to PCR product• 100% Sample Recovery• Scalable for different reaction sizes• No interference on downstream applications• Easy to automate

Degrades excess primers and dNTPs for purification of PCR products prior to:

• Sequencing• Genotyping• Cloning• SNP Analysis

PCR product

Cleaned upPCR product

dNTPs

Incubate at 37°Cfor 5 minutes

Inactivate at 80°Cfor 10 minutes

+ Rapid PCR Cleanup Enzyme Set (1 µl of each enzyme per 5 µl of PCR product)

Primer

PCR products contain residual primer and dNTPs that handicap downstream sequencing. Rapid PCR Cleanup Enzyme Set degrades excess primers and dNTPs. Add 1 µl of each enzyme (Exonuclease I and rSAP) to 5 µl of PCR product (scalable) and incubate for only 5 minutes at 37°C. The reaction is stopped by enzyme inactivation at 80°C for 10 minutes. The temperature regime can be performed in a thermocycler for maximum convenience.

Rapid PCR Cleanup Enzyme Set

Efficient removal without loss of PCR product.

Agarose gel showing three different PCR products before and after Rapid PCR Cleanup Enzyme Set treatment. No loss of PCR product was detected, irrespective of amplicon size.

PRODUCT NEB # SIZE PRICE

Rapid PCR Cleanup Enzyme Set

E2622V 20 rxns £16

E2622S 100 rxns £67

E2622L 500 rxns £268

E2622X 2000 rxns £940

E2622E 5000 rxns £1,999

PCR cleanup is important prior to sequencing. Panel A shows a sample that has undergone Rapid PCR Cleanup Enzyme Set treatment. Panel B shows a sample without this treatment. Rapid PCR Cleanup Enzyme Set treatment resulted in significant improvement in overall sequence quality.

NEW

featured product

Quick-Load Purple DNA Ladders

04

dna assembly

NEBuilder HiFi DNA Assembly was developed to improve the efficiency and accuracy of DNA assembly. This method allows for seamless assembly of multiple DNA fragments, regardless of fragment length or end compatibility. This method has been used to assemble either single-stranded oligonucleotides or different sizes of DNA fragments with varied overlaps (15–80 bp). It has utility for the synthetic biology community, as well as those interested in one-step cloning of multiple fragments due to its ease of use, flexibility and simple master-mix format. The reaction includes different enzymes that work together in the same buffer (see the workflow figure below):

• The exonuclease creates single-stranded 3´ overhangs that facilitate the annealing of fragments that share complementarity at one end (the overlap region)

• The polymerase fills in gaps within each annealed fragment

• The DNA ligase seals nicks in the assembled DNA

The end result is a double-stranded fully sealed DNA molecule that can serve as template for PCR, RCA or a variety of other molecular biology applications, including direct transformation of E coli. NEBuilder HiFi DNA Assembly Cloning Kit is supplied with NEB 5-alpha High Efficiency Competent E.coli.

NEBuilder HiFi DNA Assembly

NEBuilder HiFi DNA Assembly Workflow

A

Add fragments to NEBuilder HiFiDNA Assembly

Master Mix.

dsDNA fragments with overlapping ends.

5´3´

5´ Exonuclease chews back 5´ ends

3´5´

3´ 5´5´ 3´

Annealing

DNA polymerase extends 3´ ends

DNA ligase seals nicks

3´5´

5´3´

3´5´

5´3´

B

Fully Assembled DNA

A + B

Incubate at 50°C for 15–60 minutes.

NEBuilder HiFi DNA Assembly

Benefits over Gibson Assembly® Master Mix

• Enjoy less screening/re-sequencing of constructs, with virtually error-free, high-fidelity assembly

• Join DNA fragments together more efficiently, even with larger fragments or low DNA inputs

• Use NEBuilder HiFi in successive rounds of assembly, because it removes 5´ and 3´ end mismatches

• Bridge two double-standed fragments with a synthetic single-stranded DNA oligo for simple and fast construction (e.g., linker insertion or gDNA libraries)• Switch from other systems easily, as NEBuilder HiFi is compatible with Gibson Assembly-designed (and other) fragments

• No licensing fee requirements on NEBuilder products from NEB

NEBuilder HiFi DNA Assembly enables virtually error-free joining of DNA fragments, even those with 5´ and 3´ end mismatches. Available with and without competent E. coli, this flexible kit enables simple and fast Seamless Cloning utilizing a new proprietary high-fidelity polymerase.

Find out why NEBuilder HiFi is the next generation of DNA assembly and cloning . . .

Benefits over Traditional Cloning

• Save time with Simple and Fast Seamless Cloning

• Use one system for both “standard- size” cloning and larger gene assembly products, up to 12 fragments.

• Move on with your workflow, because DNA can be used immediately for transformation or as template for PCR or RCA.

• Adapts easily for multiple DNA manipulations, including site-directed mutagenesis.

NEW

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NEBuilder HiFi DNA Assembly Master Mix

Gibson Assembly Master Mix

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NEBuilder HiFi DNA Assembly Master Mix offers improved efficiency and accuracy over Gibson Assembly with different numbers of fragments.

NEBuilder HiFi DNA Assembly Master Mix offers improved efficiency and accuracy over Gibson Assembly with lower amounts of DNA by increasing overlaps.

Reactions were set up in a 2- and 6-fragment assembly reaction according to recommended reaction conditions.

Reactions were set up in a 4-fragment assembly reaction according to recommended reaction conditions. Amount of DNA and size of overlap is shown.

Total fragments assembled 632222 2 2

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Vector size 3.3 kb7 kb2.1 kb7.7 kb7.7 kb5.4 kb 7.7 kb 7.7 kb

Total size (kb) 8.37.75.57.97.98.4 7.9 7.9

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NEBuilder HiFi DNA Master Mix offers improved fidelity over Gibson Assembly Master Mix.

Fidelity of assembled products was compared between NEBuilder HiFi DNA Assembly Master Mix (NEB #E2621) and Gibson Assembly Master Mix (NEB #E2611). Experiments were performed using various fragment and vector sizes following suggested protocols. Experiments B through E vary because sequences of fragments are different. Experiments F and H were performed with fragments containing 3´ end mismatches.

Ordering Information

PRODUCT NEB # SIZE

NEBuilder HiFi DNA Assembly Cloning Kit E5520S 10 reactions

NEBuilder HiFi DNA Assembly Master MixE2621SE2621LE2621X

10 reactions150 reactions250 reactions

Kit Components supplied with NEBuilder HiFi DNA Assembly Cloning KitNEBuilder High-Fidelity Master MixNEB 5-alpha Competent E. coli (High Efficiency)SOC Outgrowth MediumNEBuilder Positive Control pUC19 Transformation Control Plasmid

Kit Components supplied with NEBuilder HiFi DNA Assembly Master MixNEBuilder High-Fidelity Master MixNEBuilder Positive Control

For UK prices scan this code or visit:www.neb.uk.com

References: 1. Engler, C. et al. (2008) PLoS ONE, 3: e3647. 2. Engler, C. et al. (2009) PLoS ONE, 4: e5553. 3. Lee, J.H. et al. (1996) Genetic Analysis: Biomolecular Engineering, 13; 139-145. 4. Padgett, K.A. and Sorge, J.A. (1996) Gene, 168, 31-35.

Golden Gate AssemblyThe efficient and seamless assembly of DNA fragments, commonly referred to as Golden Gate assembly (1,2), has its origins in 1996 when, for the first time, it was shown that multiple inserts could be assembled into a vector backbone using only the sequential (3) or simultaneous (4) activities of a single type IIS restriction enzyme and T4 DNA ligase. This method can be accomplished using Type IIS restriction enzymes, such as BsaI, and can also be used for the cloning of single inserts. The method is efficient and can be completed in one tube in as little as 5 minutes for single inserts, or can utilize cycling steps for multiple inserts. Golden Gate Assembly has been widely used in the construction of TALENs for in vivo gene editing, among other applications.

In its simplest form, Golden Gate Assembly requires a Type IIS recognition site, in this case, BsaI (GGTCTC), added to both ends of a dsDNA fragment. After digestion, these sites are left behind, with each fragment bearing the designed 4-base overhangs that direct the assembly.

Golden Gate Workflow

+Single-tube reaction• BsaI• DNA ligase

PCR-linearized

vector PCR-amplifiedfragments

AP3

P4

GGTCTCNNNNNCCAGAGNNNNN

5´ 3´

BsaI

NNNNNGAGACCNNNNNCTCTGG3´ 5´

P1

P2 BP5

P6

B

A

PCR amplification of vector and fragments

BsaI

Advantages of Golden Gate Assembly• Seamless cloning – no scar remains

following assembly

• Ordered assembly of multiple fragments in a single reaction

• Efficient with regions of high GC content and areas of repeats

• Compatible with a broad range of fragment sizes (< 100 bp to > 15 kb)

• Free tool available at GoldenGate.neb.com

New England Biolabs supplies reagents for use in Golden Gate Assembly, including restriction enzymes and ligases. Our new NEB Golden Gate Assembly Mix utilizes two simultaneous enzymatic activities in a single reaction, specifically digestion with BsaI and ligation with T4 DNA Ligase.

NEB Golden Gate Assembly Tool

dna assembly

06

07

NEB Golden Gate Assembly Mix offers improved assembly

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6,560

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1,663 1,636

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1,120794

1 1

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4,370

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826

Invitrogen GeneArt® Type IIs Assembly Kit, BsaI

NEB Golden Gate Assembly Mix (NEB #E1600)

Num

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NEB Golden Gate Assembly Mix E1600S 15 reactions £199COMPANION PRODUCTS

BsaI R0535S/L 1,000/5,000 units £54/£218BsaI-HF R3535S/L 1,000/5,000 units £54/£218BbsI R0539S/L 300/1,500 units £54/£218BsmBI R0580S/L 200/1,000 units £56/£225T4 DNA Ligase M0202S/L/T/M 20,000/100,000 units £52/£52/£208/£208NEB 5-alpha Competent E. coli (High Efficiency)

C2987H/I/P 20 x 0.05 ml/6 x 0.2 ml/ 1 x 96 well plate

£155/£130/£396

NEB 10-beta Competent E. coli (High Efficiency)

C3019H/I 20 x 0.05 ml/6 x 0.2 ml £168/£139


Assembly reactions were set up using either precloned inserts or PCR amplicons directly. Reaction conditions were set up according to manufacturer, and are shown above. Two separate experiments are shown for each reaction type.

FAQ Highlights

FAQ: What affects the efficiency of Golden Gate Assembly?

Single insert cloning is more efficient than multiple insert cloning. Assembly efficiency decreases as the number of fragments increases. The presence of repetitive sequences in an insert will also decrease efficiency. For inserts < 250 bp or > 3 kb, precloning will increase efficiency. Lastly, the normal restrictions on overall plasmid size to allow stable maintenance in E. coli apply to Golden Gate Assemblies. Efficiencies are highest with assembled product plasmid constructs < 12 kb. Larger assemblies can be made but will require larger numbers of colonies to be screened for the correct full length assembled products.

FAQ: Why is there a 55°C, 5 min heat step at the end of the assembly reaction?

The final incubation step at 55°C allows BsaI to perform optimally, as opposed to the 37°C temperature used in assembly reactions which allows stability of the DNA Ligase. Digesting any plasmid still present in the assembly reactions reduces background.

FAQ: How can I minimize PCR-generated errors in my amplicon inserts? Use a high-fidelity DNA polymerase and avoid over-amplification. We recommend the use of Q5® High-Fidelity DNA Polymerase formulations for maximal high-fidelity (NEB #M0491, #M0493), which is also available in Master Mix format (NEB #M0492, #M0494). Also, use the minimum number of cycles required to generate the amount of DNA required for assembly; this is usually 20 cycles or fewer.

NEW

07

08

library preparation

NEBNext Q5 Hot Start HiFi PCR Master MixThe NEBNext Q5 Hot Start HiFi PCR Master Mix is a new hot start formulation of Q5 High-Fidelity DNA Polymerase, specifically optimized for improved amplification of next-generation sequencing (NGS) libraries. This new formulation enables high yields, excellent GC coverage and ultra-high fidelity. In addition, compatibility with magnetic beads used in NGS workflows is enhanced, as is tolerance to over-cycling. This combination makes the NEBNext Q5 Hot Start HiFi PCR Master Mix an ideal choice for your library construction.

The convenient 2X master mix contains dNTPs, Mg2+ and a proprietary buffer, and requires only the addition of primers and DNA template for robust amplification. Inclusion of the hot start aptamer allows convenient room temperature reaction setup.

FidelityHigh-fidelity amplification of NGS libraries, with minimal GC bias, is critical for generation of high quality libraries, especially when input amounts are limiting. The polymerase component of the NEBNext Q5 Hot Start HiFi PCR Master Mix, Q5 High-Fidelity DNA Polymerase, is a novel thermostable DNA polymerase that possesses 3´-5´ exonuclease activity, and is fused to a processivity-enhancing Sso7d domain. Q5 also has an ultra-low error rate (> 100-fold lower than that of Taq DNA Polymerase and ~12-fold lower than that of Pyrococcus furiosus (Pfu) DNA Polymerase). The buffer component of this new master mix formulation has been optimized for robust high-fidelity amplification, even with GC-rich amplicons.

Advantages• Minimizes GC bias

• Hot start (aptamer based)

• High yields

• Ultra high fidelity amplification

• Compatible with AMPure® beads, and other magnetic beads used in NGS workflows

• Improved tolerance to over-cycling

Application• Amplification of libraries for next

generation sequencing

NEBNext Q5 Hot Start HiFi PCR Master Mix minimizes GC bias

Libraries of human (IMR-90) or E. coli K12 genomic DNA were unamplified (“PCR-free”) or amplified using NEBNext Q5 Hot Start HiFi PCR Master Mix, and sequenced on an Illumina® MiSeq®. GC bias plots were generated, with %GC content of 100 bp windows on the X axis. Normal-ized coverage is indicated by the blue circles, %GC of the reference sequence indicated by the red lines and base quality at %GC indicated by the green line. Similar GC bias was observed for the libraries amplified with NEBNext Hot Start HiFi PCR Master Mix compared to PCR-free libraries.



NEBNext Q5 Hot Start HiFi PCR Master MixM0543SM0543L

5 rxns250 rxns

£82£328

E. coli

HumanPCR-free

PCR-free

NEBNext Q5 Hot Start HiFi PCR Master Mix-amplified

NEBNext Q5 Hot Start HiFi PCR Master Mix-amplified

For licensing information, visit www.neb.com.

NEW

For Research Use Only. Not For Use in Diagnostic Procedures.

09

NEBNext FFPE DNA Repair MixArchiving of clinical materials as Formalin-Fixed, Paraffin-Embedded (FFPE) samples is a common practice. However, the methods used for fixation and storage significantly damage and compromise the quality of nucleic acids from these samples. As a result, it can be challenging to obtain useful information, including high-quality sequence data, especially when sample amounts are limited. The NEBNext FFPE DNA Repair Mix is a cocktail of enzymes formulated to repair DNA, and specifically optimized and validated for repair of FFPE DNA samples. Incorporation of the FFPE DNA Repair Mix into Next Generation Sequencing (NGS) workflows can increase yields and overall library success rates, without introduction of bias.

Benefits• Construct high-quality NGS libraries from

FFPE DNA samples

• Use before library prep for any NGS platform

• No alteration of DNA sequence

• Rely on NEB’s NGS validation process for FFPE DNA library prep

An example of Agilent Bioanalyzer® traces of libraries prepared from stomach tumor FFPE DNA that was treated with the FFPE DNA Repair Mix, or was untreated, before library construction.Yield improvements of 101% to 458% have been observed.

Effect of FFPE DNA Repair Mix on library yields

– FFPE DNA Repair+ FFPE DNA Repair

For more information or to request a sample, contact [email protected] licensing information, visit www.neb.com.


Effect of DNA Repair on Sequence Quality from FFPE Samples

Genomic DNA was extracted using standard DNA purification. Each DNA sample was treated with the NEBNext FFPE DNA Repair Mix, or was untreated, before library construction and Illumina sequencing.Panel A: The effect of DNA repair on the percentage of mappable reads. Reads were mapped to the human genome (hg19) using BWA (1). Panel B: The effect of DNA repair on the percentage of reads that were properly paired from a paired-end sequencing protocol.

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1. Li H. and Durbin R. (2009) Fast and accurate short read alignment with

Burrows-Wheeler Transform. Bioinformatics, 25:1754-60.


NEBNext FFPE DNA Repair MixM6630SM6630L

24 rxns96 rxns

£140£489

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library preparation

For Research Use Only. Not For Use in Diagnostic Procedures.

CHROMATIN IP

The success or failure of a ChIP experiment is highly dependent on the integrity of the chromatin, the quality of the epitope, and the specificity of the antibody. Just as important is the inclusion of a control antibody that binds at the locus of interest and allows the investigator to confidently assess results. These components must be optimized to work together, especially when the target interaction is a low abundance, low stability event.

High abundance, very stable protein-DNA interactions like those between histones and DNA, occur frequently enough that they may still be detected even if the integrity of the DNA or protein epitopes has been compromised, or if the signal to noise ratio of the antibody is low.

Low abundance, less stable interactions such as the bind-ing of polycomb group proteins (e.g., Ezh2) to specific genes, may fall under the detection threshold if the protocol fails to safe-guard the integrity of the protein and the DNA, or if it relies on an antibody that is not highly specific to the target of interest.

EZH2EED

RBAP48

PcG

SUZ12

EZH2EED

RBAP48

PcG

SUZ12

Nucleosome

Ezh2 (D2C9) XP® Rabbit mAb #5246

Normal Rabbit IgG #2729

% o

f Tot

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put

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Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733

GAPDHRPL30HoxA1HoxA2

High AbundanceHigh Stability

Low AbundanceLow Stability

High Enrichment Low Enrichment

Optimized Reagents MatterSimpleChIP® Plus Chromatin IP Kits and ChIP-validated Antibodies

APPLICATION FOCUS

SimpleChIP ®Enzymatic Chromatin IP Kits and

ChIP-validated Antibodies

Request a SimpleChip Brochure

For more information on CST’s

ChIP Kits and protocols, including

data on enzymatic

digestion versus

sonication please

use the Literature

Request Form at

www.neb.uk.com

CHROMATIN IMMUNOPRECIPITATION

Antibody Specificity: CST™ ChIP-Validated Antibodies

Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb #12041Antibodies that non-specifically bind unintended targets increase the background noise, making it more difficult to detect low abundance interactions.

CST offers antibodies that have been validated to work in ChIP applications, using the same rigorous standards we apply to all our antibodies.

Please visit www.cellsignal.com/cstchipab for a full list of ChIP validated antibodies and primer pairs.

Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb #12041 showed efficient target enrichment only when the cells were treated with dexamethasone, indicating the antibody is highly specific for the target of interest.Western blot analysis of extracts from various cell lines using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb #12041(upper).

A549 cells were cultured in media with 5% charcoal-stripped FBS for 3 d and then either untreated (left panel) or dexamethasone-treated (100 nM, 1 hr; right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 cells and 10 µl of Glucocorticoid Receptor (D6H2L) Rabbit mAb #12041or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human SLC19A2 Promoter Primers #7681, human MT2A promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as percent of the total input chromatin. (lower)

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A. U-251 MGB. NCI-H295RC. OVCAR8D. DU 145E. 786-OF. A549G. BT-549

H. MCF7I. MOLT-4J. CCRF-CEMK. HeLaL. L-929M. Raw 264.7N. COS-7

A B C D E F G H I J K L M N

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Untreated Dexamethasone Treated

These kits contain all reagents necessary to perform enzymatic digestion-based chromatin immunoprecipitation (ChIP) experiments quickly and easily, as well as positive and negative controls that allow you to be confident in your results. These kits are available with either Protein G agarose or Protein G magnetic beads and contain all buffers and reagents needed to perform up to 30 ChIP assays.

SimpleChIP® Plus Chromatin IP Kits from CST detect endogenous protein-DNA interactions in cultured cells and tissue samples.

Chromatin IntegrityEnzymatic digestion gently fragments the chromatin, protecting the integrity of the protein and the DNA.

Assay ReliabilityThe Histone H3 antibody is a universal control for tracking assay efficiency and reagent performance.

Antibody SpecificityCST ChIP-validated antibodies are rigorously tested and validated, ensuring they will specifically bind to their intended target.

Each kit is designed to optimize:

1 2 3

Cat.# Name Application Reactivity#9004 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads) ChIP H, M, R, Mk

#9005 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) ChIP H, M, R, Mk

SimpleChIP® Plus Enzymatic Chromatin IP Kits

www.cellsignal.com

IMMUNE CHECKPOINTS

Proven specificity & sensitivity. Results you can count on.Antibodies for PD-L1, B7-H3, B7-H4, Phospho-SLP-76 (Ser376), Phospho-Stat3 (Tyr705), and more from CST.

For Research Use Only. Not For Use In Diagnostic Procedures. © 2015 Cell Signaling Technology, Inc. Cell Signaling Technology, CST, E1L3N and XP are trademarks of Cell Signaling Technology, Inc.

www.cellsignal.com/cancerimmunoVisit our website to request our Tumor Immunology Poster and for additional validation and competitor comparison data.

CST development scientist optimizing IHC protocols

PIVOTAL TARGETS

CANCER IMMUNOLOGY

PD-L1 (E1L3N®) XP® Rabbit mAb #13684:

IHC analysis of paraffin-embedded human lung

carcinoma using #13684.

Phospho-SLP-76 (Ser376) (D9D6E) Rabbit mAb #14745: Flow cytometric analysis of human PBMCs stimulated with anti-human CD3/CD28 using #14745.

expressions - new england biolabs (uk) ltdavailable with and without competent e. coli, this...

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