Strep

Strep

Cloning Cell culture ScreeningScale-up

purifi cationStructural

& functional studies

Lead design

E. coli.

GST-tagged proteins

Schistosoma japonicumE. coli

pGEX-1 T (27-4805-01)

pGEX-6P-1 (27-4597-01)

EcoR I Sma I Sal I Xho I Not IBamH I

PreScission Protease

Leu Glu Val Leu Phe Gln Gly Pro Leu Gly Ser Pro Glu Phe Pro Gly Arg Leu Glu Arg Pro HisCTG GAA GTT CTG TTC CAG GGG CCC CTG GGA TCC CCG GAA TTC CCG GGT CGA CTC GAG CGG CCG CAT

pGEX-6P-2 (27-4598-01)

BamH I

PreScission Protease

Leu Glu Val Leu Phe Gln Gly Pro Leu Gly Ser Pro Gly Ile Pro Gly Ser Thr Arg Ala Ala Ala SerCTG GAA GTT CTG TTC CAG GGG CCC CTG GGA TCC CCA GGA ATT CCC GGG TCG ACT CGA GCG GCC GCA TCG

EcoR I Sma I Sal I Xho I Not I

pGEX-6P-3 (27-4599-01)

BamH I

PreScission Protease

Leu Glu Val Leu Phe Gln Gly Pro Leu Gly Ser Pro Asn Ser Arg Val Asp Ser Ser Gly ArgCTG GAA GTT CTG TTC CAG GGG CCC CTG GGA TCC CCG AAT TCC CGG GTC GAC TCG AGC GGC CGC

EcoR I Sma I Sal I Xho I Not I

BamH I EcoR I Sma I Sal I Xho I Not I

BamH I EcoR I Sma I Sal I Xho I Not I

BamH I EcoR I Sma I Sal I Xho I Not I

BamH I EcoR I Sma I Sal I Xho I Not I

BamH I EcoR I Sma I Sal I Xho I Not I

BamH I EcoR I Sma I Sal I Xho I Not I

EcoR I

CTG GTT CCG CGT GGA TCC CCG GAA TTC ATC GTG ACT GAC TGA CGA

BamH I

Leu Val Pro Arg Gly Ser Pro Glu Phe Ile Val Thr Asp

Thrombin Protease

Stop codons

pGEX ~4900 bp

pBR322 ori

Bal I

BspM I

Ptac

c a l

I q

Nar I

EcoR V

BssH II

BstE II Mlu I

Apa I

Tth111 I Aat II

Pst I

p4.5 AlwN I

pSj10 Bam7Stop7

pGEX-4T-2 (27-4581-01)

pGEX-5X-1 (27-4584-01)

pGEX-5X-2 (27-4585-01)

pGEX-5X-3 (27-4586-01)

pGEX-4T-1 (27-4580-01)

pGEX-4T-3 (27-4583-01)

pGEX-3X (27-4803-01)

pGEX-2TK (27-4587-01)

Leu Val Pro Arg Gly Ser Pro Gly Ile Pro Gly Ser Thr Arg Ala Ala Ala SerCTG GTT CCG CGT GGA TCC CCA GGA ATT CCC GGG TCG ACT CGA GCG GCC GCA TCG TGA

Stop codon

Ile Glu Gly Arg Gly Ile Pro Glu Phe Pro Gly Arg Leu Glu Arg Pro His Arg AspATC GAA GGT CGT GGG ATC CCC GAA TTC CCG GGT CGA CTC GAG CGG CCG CAT CGT GAC TGA

Stop codons

Ile Glu Gly Arg Gly Ile Pro Gly Ile Pro Gly Ser Thr Arg Ala Ala Ala SerATC GAA GGT CGT GGG ATC CCC GGA ATT CCC GGG TCG ACT CGA GCG GCC GCA TCG TGA

Stop codon

Ile Glu Gly Arg Gly Ile Pro Arg Asn Ser Arg Val Asp Ser Ser Gly Arg Ile Val Thr AspATC GAA GGT CGT GGG ATC CCC AGG AAT TCC CGG GTC GAC TCG AGC GGC CGC ATC GTG ACT GAC TGA

Stop codons

Leu Val Pro Arg Gly Ser Pro Glu Phe Pro Gly Arg Leu Glu Arg Pro His Arg AspCTG GTT CCG CGT GGA TCC CCG GAA TTC CCG GGT CGA CTC GAG CGG CCG CAT CGT GAC TGA

Stop codons

Leu Val Pro Arg Gly Ser Pro Asn Ser Arg Val Asp Ser Ser Gly Arg Ile Val Thr AspCTG GTT CCG CGT GGA TCC CCG AAT TCC CGG GTC GAC TCG AGC GGC CGC ATC GTG ACT GAC TGA

Stop codons

ATC GAA GGT CGT GGG ATC CCC GGG AAT TCA TCG TGA CTG ACT GACIle Glu Gly Arg Gly Ile Pro Gly Asn Ser Ser

Stop codons

Leu Val Pro Arg Gly Ser Arg Arg Ala Ser Val

Kinase

CTG GTT CCG CGT GGA TCT CGT CGT GCA TCT GTT GGA TCC CCG GGA ATT CAT CGT GAC TGAStop codons

Thrombin Protease

Thrombin Protease

Thrombin Protease

Thrombin Protease

Factor Xa Protease

Factor Xa Protease

Factor Xa Protease

Factor Xa Protease

EcoR IBamH I Sma I

EcoR IBamH I Sma I

pGEX-2T (27-4801-01)

CTG GTT CCG CGT GGA TCC CCG GGA ATT CAT CGT GAC TGA CTG ACGLeu Val Pro Arg Gly Ser Pro Gly Ile His Arg Asp

Stop codons EcoR I

Thrombin Protease

BamH I Sma I

glutathione S-transferase

Amp r

GST

-tag

ged

prot

eins

Glutathione Sepharose

* Purification modules also contain buffers

GST-tagged proteins

Products featured: GST MultiTrap FF and GST MultiTrap 4B, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

E. coli

96-well filter plates:

Sample: E. coli

Sample preparation:

Binding buffer:

Elution buffer:

Elution method:

SDS-PAGE

500

450

400

350

300

250

2000 20 40 60 80 100 120 140

Yield GST-hippocalcin, GST MultiTrap FFYield GST-hippocalcin, GST MultiTrap 4B

Incubation time (min)

Aver

age

yiel

d of

elu

ted

prot

ein

(μg)

97

66

45

30

2014

(Mr × 103)

GST-hippocalcin

LMW

mar

kers

Star

t mat

eria

lFl

owth

roug

hEl

uate

Flow

thro

ugh

Elua

teFl

owth

roug

hEl

uate

Flow

thro

ugh

Elua

teFl

owth

roug

hEl

uate

Flow

thro

ugh

Elua

te

120 min 60 min 30 min 10 min 3 min 0 min

Summary

GST

-tag

ged

prot

eins

Products featured: GST SpinTrap Purification Module, GST Detection Module

96-well plate

GST-tagged proteins

Column:

Sample: β

Binding buffer:

Elution buffer:

SDS-PAGE

DDM OG CHAPS Tween 20Triton X-100 Sarcosyl

GST-ecoKch94

67

43

(Mr × 103)

LMW

mar

kers

Flow

thro

ugh

Elua

te fr

om G

ST S

pinT

rap

Flow

thro

ugh

Elua

te fr

om G

ST S

pinT

rap

Flow

thro

ugh

Elua

te fr

om G

ST S

pinT

rap

Flow

thro

ugh

Elua

te fr

om G

ST S

pinT

rap

Flow

thro

ugh

Elua

te fr

om G

ST S

pinT

rap

Flow

thro

ugh

Elua

te fr

om G

ST S

pinT

rap

0.8

0.6

0.4

0.2

0

Solubilized materialFlowthroughEluate

Activ

ity (A

340/

min

*ml)

DDM OGCHAPS

Tween 20

Triton X-100

Sarcosyl

Summary

Acknowledgements:

GST

-tag

ged

prot

eins

Products featured: GSTrap FF, ÄKTAexplorer, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Column:

Sample: E. coli

Binding buffer:

Elution buffer:

Flow:

System:

SDS-PAGE

0

20

40

60

80

100

%Elutionbuffer

0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

ml

Wash

Elutionbuffer

2.7 mgpure GST-taggedprotein

5.0 10.0 15.0 20.0

A280

97

66

45

30

2014

(Mr × 103)

LMW

mar

kers

Star

t mat

eria

lEl

uted

frac

tion

Summary

GST-tagged proteins

Summary

Products featured: GSTrap 4B, HiLoad 16/60 Superdex 200 pg, ÄKTAxpress, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Columns:

Sample: E. coli

Sample volume:

Binding buffer (AC):

Elution buffer (AC):

Buffer (GF):

Flow rates:

System:

SDS-PAGE

500

0

1000

1500

mAU

00205 150100

AC GF

ml

A280

Large aggregates ofGST-hippocalcin

Dimers of GST-hippocalcin

150140130120110 ml

50

0

100

150

200

250

mAUA280

9766

45

30

20

14

(Mr × 103)

GST-hippocalcin

LMW

mar

kers

Star

t mat

eria

lFl

owth

roug

h (A

C)

Elut

ed fr

actio

ns o

f dim

er (G

F)

GST

-tag

ged

prot

eins

Products featured: pGEX-2T, GSTrap FF, GSTPrep FF 16/10, ÄKTAexplorer, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Columns:

Sample: E. coli

Sample volumes:

Binding buffer:

Elution buffer:

Flow rates:

System:

SDS-PAGE

Summary

mAU

1200

1000

800

600

400

200

0

A 280 Elution buffer

100

80

60

40

20

0

%B

0

Wash

Elutionbuffer

5.5 mg

10 20 30 ml

1400

1200

1000

800

600

400

200

0

A 280

100

80

60

40

20

0

%B

Wash

Elutionbuffer

20.9 mgmAU

0 50 100 150 ml

Elution bufferElution buffer

A280

mAU 1400

1200

1000

800

600

400

200

0

Elution buffer

100

80

60

40

20

0

%B

Wash

111.0 mg

0 100 200 300 400 500 600 ml

GST-DemA

9766

45

30

20

14

(Mr × 103)

LMW

mar

kers

Star

t mat

eria

l

Star

t mat

eria

l

Flow

thro

ugh

Elut

ed fr

actio

ns

Flow

thro

ugh

Elut

ed fr

actio

nsSt

art m

ater

ial

Flow

thro

ugh

Elut

ed fr

actio

ns

GSTrap FF 1 ml GSTrap FF 5 ml GSTPrep FF 16/10

GSTrap FF 1 ml GSTrap FF 5 ml GSTPrep FF 16/10

Cleavage site

Glutathione

GST

Recombinant protein

Sepharose

Cleavage enzymesFactor Xa protease:

Thrombin protease:

PreScission Protease:

GST-tagged proteins

GST

-tag

ged

prot

eins

Products featured: GSTrap FF, PreScission Protease, ÄKTAxpress

Summary

0

10

20

30

40

50

60

70

80

90

100

10 units PreScission/mg protein20 units PreScission/mg protein40 units PreScission/mg protein

2 4 6 8 10 12 14Incubation time (h)

Cle

aved

pro

tein

(%

)

0

GST-tagged proteins

Products featured: GSTrap FF, Thrombin protease, ÄKTAexplorer, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

SDS-PAGE

Summary

Column:

Sample: E. coli

Binding buffer:

Elution buffer:

Flow rate:

System:

0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

5.0 10.0 15.0 min

WashIncubation

16 hroom temp.

Wash

A280

Targetprotein

FreeGST

2.0 4.0 6.0 8.0 10.0 12.0 min

0

20

40

60

80

100

%Elutionbuffer

97

66

45

30

20

14

(Mr × 103)

LMW

mar

kers

Star

t mat

eria

l

GST-

tagg

ed p

rote

in, 1

ml c

olum

n

GST-

tagg

ed p

rote

in, 5

ml c

olum

n

GST-

free

targ

et p

rote

in, 1

6 h

clea

vage

Clea

ved

GST

tag

Thro

mbi

n pr

otea

se (2

0 U/

ml)

LMW

mar

kers

Thrombin protease cleavage,

GSTrap FF 1 ml

Target protein

GST

-tag

ged

prot

eins

Products featured: pGEX-6P, GSTrap HP, PreScission Protease, HiLoad 16/60 Superdex 75 pg, ÄKTAxpress, ExcelGel SDS gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Summary

Columns:

Sample: α

Binding/cleavage buffer (AC):

Elution buffer (AC):

Buffer (GF):

System:

SDS-PAGE

AC GF

0

500

1000

1500

2000

mAU

200 250 300

46 mg

Cleaved protein

Regeneration

A 280

ml

97

66

45

30

20

14

(Mr × 103)

LMW

mar

kers

Star

t mat

eria

lFl

owth

roug

h

Purif

ied,

clea

ved

GST-

pur

Uncl

eave

d GS

T-pu

r

pur

GST-tagged proteins

4 5 6 7 8 9 10 11 12 1 2 3

Control Anti-luciferase detection

Columns

Products featured: pGEX-6P-1, GST 96-well Detection Module

E. coli

Summary

GST

-tag

ged

prot

eins

SDS-PAGE

9766

45

30

20

14

(Mr × 103)

Soni

cate

, E. c

oli T

G1

Soni

cate

, KL4

5 ce

lls (D

naK

over

expr

essin

g)

Soni

cate

, E. c

oli +

pGE

X-5X

-1

Soni

cate

, E. c

oli +

pGE

X-5X

-Luc

exp

ress

ing

GST-

luci

fera

se

Soni

cate

, E. c

oli +

pGE

X-4T

-E7

expr

essin

g GS

T-E7

Purif

ied

GST

Pres

tain

ed M

W m

arke

r

Products featured: Anti-GST Antibody, Amersham Hybond ECL, Amersham Hyperfilm ECL, Amersham GST Western Blotting Detection Kit

E. coli E. coli

E. coli

HRP

Summary

Product Quantity Code No.

Protein expression

λ

All vectors include BL21

Purification

Detection

Tag cleavage

Related products

Product Quantity Code No.

† available by specific customer order

For more information on equipment for chromatography and/or electrophoresis, please visit www.gelifesciences.com

GST-tagged proteins

Histidine-tagged proteins

Ni Sepharose

His

tidin

e-ta

gged

pro

tein

s

Products featured: His MultiTrap HP

Summary

96-well filter plate:

Samples: E. coli

E. coli

Equilibration buffer:

Wash buffer:

Elution buffer:

Liquid handling station:

Vacuum station:

100

75

50

37

2520

15

10

1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6M M M M

Clarified bacterial extracts

Elution, row 4

Elution,row 1

Elution, row 2

Elution, row 3

Elution, row 5

Elution, row 6

Elution, row 7

(Mr × 103)

M B P B P B P B P B P B P B P B P B P B P B P B P B

B : Blank

P : Protein purification(37 kDa)

B : Blank

P : Protein purification(37 kDa)

B : Blank

P : Protein purification(37 kDa)

150

10075

50

37

2520

15

10

(Mr × 103)

Acknowledgements:

Histidine-tagged proteins

Products featured: His SpinTrap, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

97

66

45

30

20

14

(Mr × 103)

APB 7-(His)6

LMW

mar

kers

Star

t mat

eria

l, di

lute

d 1:

10

5 m

M im

idaz

ole

durin

g bi

ndin

g, d

ilute

d 1:

2

50 m

M im

idaz

ole

durin

g bi

ndin

g, d

ilute

d 1:

2

100

mM

imid

azol

e du

ring

bind

ing,

dilu

ted

1:2

200

mM

imid

azol

e du

ring

bind

ing,

dilu

ted

1:2

Eluted pool

Column:

Sample: E.coli

Binding/wash buffer:

Elution buffer:

Summary

SDS-PAGE

His

tidin

e-ta

gged

pro

tein

s

Summary

Products featured: His GraviTrap Kit, PhastSystem, PhastGel Gradient 10–15, Amersham Hybond ECL, Anti-His Antibody, ECL Mouse IgG, HRP-linked Whole Ab, Amersham High-Range Rainbow Molecular Weight Markers

E. coli

High

mol

ecul

ar w

eigh

t mar

kers

Star

t mat

eria

l, di

lute

d 1:

20

Flow

thro

ugh,

dilu

ted

1:20

Elua

te 1

Elua

te 2

Nega

tive

cont

rol (

untr

ansf

orm

ed JM

109)

220

97

66

45

20

14

30

(Mr × 103)

High

mol

ecul

ar w

eigh

t mar

kers

Star

t mat

eria

l, di

lute

d 1:

20

Flow

thro

ugh,

dilu

ted

1:20

Elua

te 1

Elua

te 2

Nega

tive

cont

rol (

untr

ansf

orm

ed JM

109)

Western blot

Column:

Sample: E.coli

Binding/wash buffer:

Elution buffer:

SDS-PAGE

Histidine-tagged proteins

Products featured: HisTrap FF crude Kit, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Summary

030252015

Wash 1 Wash 2 Wash 3Elution 1

10500

0.5

1.0

1.5

2.0

2.5

3.0

100

200

300

400

500

Imidazole concentration

(mM)

Elution volume (ml)

A280

Elution 2

Elution 3

(Mr × 103)

9766

45

30

20

14

MBP-(His)6

LMW

mar

kers

Uncl

arifi

ed sa

mpl

e, d

ilute

d 1:

10

Was

h 5

ml,

10 m

M im

idaz

ole

Was

h 1,

10

mM

imid

azol

e

Was

h 2,

30

mM

imid

azol

e

Was

h 3,

50

mM

imid

azol

e

Elut

ion

1, 1

00 m

M im

idaz

ole

Elut

ion

2, 3

00 m

M im

idaz

ole

E

lutio

n 3,

500

mM

imid

azol

e

Column:

Sample: E. coli

Flow rate:

Buffers:

Fraction size:

Equipment:

SDS-PAGE

His

tidin

e-ta

gged

pro

tein

s

Summary

Products featured: HisTrap HP, ÄKTAprime plus

Sample: E. coli

Column:

Binding buffer:

Elution buffer:

System:

5 10 15 20 25 30 35 min

2.5

2.0

1.5

1.0

0.5

0

100

80

60

40

20

0

A280Elution

Buffer %

Histidine-tagged proteins

Pichia pastoris Products featured: HisTrap FF crude, ÄKTAexplorer, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Pichia pastoris

Summary

Column:

Sample: Saccharo-myces cerevisiae P. pastoris

Binding buffer:

Elution buffer:

Flow rate:

System:

mAU

5000

4000

3000

2000

1000

0

100

50

0

0 100 200 300 400 ml

Elution Buffer %

A280

(Mr × 103)

9766

45

30

2014

(His)6-YNR064c

LMW

mar

kers

Star

t mat

eria

l

Elut

ed p

ool,

dilu

ted

1:2

Elut

ed p

ool,

dilu

ted

1:4

SDS-PAGE

His

tidin

e-ta

gged

pro

tein

s

Products featured: HisTrap HP, ÄKTAexplorer, Biacore System

E. coli.

E. coli

Summary

500

400

300

200

100

0

91.5 92 93

pool

94 95 ml92.5 93.5 94.5

Elution buffer %100

80

60

40

20

0

500

400

300

200

100

0

0 20 40 60 80 ml

Elution buffer %mAU

100

80

60

40

20

0

1

42

3 5

Pooled fraction from the purification of refolded scFv 57P antibody fragment

100

A280

A280

mAU

Sample:

Column:

Solubilizing buffer:

Denatured binding buffer:

Refolding buffer:

Native binding buffer:

Native elution buffer:

Flow rate:

System:

5000

0

10 000

15 000

20 000

-5000

Resp

onse

sig

nal (

RU)

Reference cellSample cellResponse difference after subtraction of reference cell

200 400 600 800 1000 1200 1400 1600s

SPR sensogram

Histidine-tagged proteins

Summary

Products featured: HisTrap HP, HiPrep 26/10 Desalting, RESOURCE Q, HiLoad 16/60 Superdex 75 pg, ÄKTAxpress, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

0

500

1000

1500

mAU

1100 1200 1300 1400 1500B10

11 mg

Cleaved protein

Regeneration

A280

ml

Elution buffer %

100

80

60

40

20

0

AC DS IEX GF

97

66

45

30

20

14

(Mr × 103)

Cleaved APC1040 (Mr 36 400)Uncleaved APC1040 (Mr 38 900)

LMW

mar

kers

Star

t mat

eria

lFl

owth

roug

h

Purif

ied,

clea

ved

APC1

040

afte

r AC–

DS–I

EX–G

F

Cont

rol,

uncl

eave

d AP

C104

0

Sample: E. coli

Columns: AC:

DS:

IEX:

GF:

Cleavage conditions:

AC binding buffer:

AC cleavage buffer:

AC elution buffer:

DS and IEX binding buffer:

IEX elution buffer:

GF buffer:

His

tidin

e-ta

gged

pro

tein

s

Products featured: HisTrap FF columns, HisPrep FF 16/10, ÄKTAexplorer, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Summary

Columns:

Sample: E. coli

Sample volumes:

Binding buffer:

Elution buffer:

Flow rates:

System:

SDS-PAGE

A280

0.0 50 100 150 ml

mAU4000350030002500200015001000500

0

33 mg

100

0

50

Elutionbuffer %

HisTrap FF 1 ml HisTrap FF 5 ml HisPrep FF 16/10

0.0 5.0 10.0 15.0 20.0 25.0 30.0 40.0 ml

mAU4000350030002500200015001000

5000

6.2 mg

A280

100

0

50

Elutionbuffer % 100

0

50

Elutionbuffer %

A280

mAU4000350030002500200015001000500

00 100 200 300 500400 600 ml

149 mg

700

97

66

45

30

20

14

(Mr × 103)

MBP-(His)6

LMW

mar

kers

Star

t mat

eria

l

Elut

ed p

ool H

isTra

p FF

1 m

l

Elut

ed p

ool H

isTra

p FF

5 m

l

Elut

ed p

ool H

isPre

p FF

16/

10

Histidine-tagged proteins

Product Quantity Code No.

Purification Detection

Related products

Product Quantity Code No.

For more information on Biacore systems and chromatography and/or electrophoresis systems, please visit www.gelifesciences.com

available by specific customer order

Strep-

Strep

Strep Strep

Strep

Strep

Strep-tag II proteins

Products featured: StrepTrap HP, HiLoad 16/60 Superdex 75 pg, ÄKTAprime plus, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

StrepStrep E. coli

AC step:Column: Sample: E. coli

Strep

Flow rate: Binding buffer:

Elution buffer:

GF step:Column: Sample: Flow rate: Buffer:System:

2000

1500

0

A280 mAU

1000

500

0 10 20 30 40 ml

40

50

30

60

70

0

20

10

0 10 20 30 40 50 60 ml

A280 mAU

SummaryStrep

(Histidine)6 tag Strep-tag IImCherry Full-length target protein

(Histidine)6 tag

Strep-tag II

mCherry fragment

mCherry fragment

PreScission MYG (85-87) TEV

PreScission

MYG (85-87) TEV

97

66

45

30

20

14

(Mr × 103)

SDS-PAGE

AC step GF step

Stre

p-ta

g II

prot

eins

Products featured: StrepTrap HP, ÄKTAxpress

StrepStrep Strep

Column:

Sample: Strep

Binding buffer:

Elution buffer:

Flow rates:

System:

188

98

6249

38

28

1714

60

30

(Mr × 103)

(His)6-Strep- tag II-protein

Mar

kers

Lysa

te

Flow

thro

ugh

Elut

ed ta

rget

pro

tein

poo

l

Summary

SDS-PAGE

Acknowledgements:

Strep-tag II proteins

Products featured: StrepTrap HP columns, XK 26/20 column, StrepTactin Sepharose High Performance, ÄKTAexplorer

StrepE. coli

Summary

0

1000

2000

3000

A280 mAU

0

1000

2000

3000

A587mAU

0.0 10.0 20.0 ml

= A280

= A587

= Elution buffer, %B

Columns:

Sample: Strep E. coli

Sample volumes:

Regeneration:

Binding buffer:

Elution buffer:

Flow rates:

System:

A280 mAU

A587mAU

0

1000

2000

3000

0

1000

2000

3000

0 50 100 150 ml

A280 mAU

A587mAU

0

1000

2000

3000

0

1000

2000

3000

0 200 400 600 700 ml

StrepTrap HP 1 ml StrepTrap HP 5 ml StrepTactin Sepharose High Performance XK 26/20

Stre

p-ta

g II

prot

eins

Purification

Related products

Product Quantity Code No.

For more information on chromatography and/or electrophoresis systems, please visit www.gelifesciences.com

MBP-tagged proteins

Products featured: MBPTrap HP, HiLoad 16/60 Superdex 200 pg, ÄKTAprime plus, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

δ

Summary

2000

1500

0

1000

500

0 10 20 30 40 ml

A280mAU

100

120

50

A280mAU

0

0 20 40 60 80 100 120 ml

AC step:Column: Sample: δ E. coli

Flow rate: Binding buffer:

Elution buffer:

GF step: Column: Sample: Flow rate: Buffer:

97

66

45

30

20

14

(Mr × 103)

MBP*2-paramyosin-

-SalLM

W m

arke

rsSa

mpl

e, d

ilute

d 1:

15

Flow

thro

ugh,

dilu

ted

1:15

, MBP

Trap

HP

MBP

Trap

HP

HiLo

ad S

uper

dex 1

6/60

200

pg

Eluted pool

SDS-PAGE

MBP

-tag

ged

prot

eins

Products featured: MBPTrap HP, Superdex 200 prep grade, XK 16/20, ÄKTAprime

Summary

1000

1500

2000

2500

500

A280

mAU

0

0 20 40 60 80 100 ml120

200

300

400

100

0

0 20 40 60 80 ml

A280

mAU

170130

9572

56

43

34

26

1117

(Mr × 103)

MBP-MCAD

Mol

ecul

ar w

eigh

t mar

kers

Star

t mat

eria

l, di

lute

d 1:

6

Flow

thro

ugh

MBP

Trap

HP,

dilu

ted

1:6

Elut

ed fr

actio

ns fr

om M

BPTr

ap H

PEl

uted

frac

tions

from

Sup

erde

x 200

pg

AC stepColumn: Sample: E. coli

Binding buffer:

Elution buffer: Flow rate: System:

GF stepColumn: Sample: Buffer:Flow rate: System:

SDS-PAGE

Acknowledgements:

MBP-tagged proteins

Products featured: MBPTrap HP, HiLoad 16/60 Superdex 200 pg, ÄKTAxpress

Summary

AC column:

Sample: E. coli

Flow rate:

Binding buffer:

Elution buffer:

GF column:

Sample:

Flow rate:

Buffer:

System:

Elution buffer%B

0

500

1000

1500

2000

A280mAU

0

20

40

60

80

100

60 80 100 120 140 160 180 mlAC GF

SDS-PAGE

Acknowledgements:

250130100

70

55

35

27

15

10

(Mr × 103)

Mol

ecul

ar

wei

ght m

arke

r Eluted fractions from gel filtration

MBP

-tag

ged

prot

eins

Products featured: MBPTrap HP columns, XK 26/20 column, Dextrin Sepharose High Performance, ÄKTAexplorer, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

β

Summary

Columns:

Sample: β E. coli

Sample volumes:

Binding buffer:

Elution buffer:

Flow rates:

System:

0

500

1000

1500

2000

2500

3000

A280mAU

0.0 5.0 10.0 15.0 20.0 ml

80

100

Elution Buffer %

60

40

20

0 0

500

1000

1500

2000

2500

3000

0 100 200 300 400 500 ml

80

100

60

40

20

0600

A280mAU

Elution Buffer %

97

66

45

30

20

14

(Mr × 103)

MBP2*- -galactosidase

LMW

mar

kers

MBP

Trap

HP

1 m

l, di

lute

d 1:

6

MBP

Trap

HP

5 m

l, di

lute

d 1:

12

XK 2

6/20

, dilu

ted

1:12

MBP

Trap

HP

1 m

l, di

lute

d 1:

3

MBP

Trap

HP

5 m

l, di

lute

d 1:

3

XK 2

6/20

, dilu

ted

1:3

Star

t mat

eria

l, di

lute

d 1:

3

FlowthroughEluted pool

MBPTrap HP 1 ml Dextrin Sepharose High Performance XK 26/20

SDS-PAGE

MBP-tagged proteins

Purification

Purification

Product Quantity Code No.

For more information on Biacore systems and chromatography and/or electrophoresis systems, please visit www.gelifesciences.com

Dual-tagged proteinsProducts featured: HisTrap HP, GSTrap 4B, ÄKTAxpress, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Summary

Sample: E.coli

Columns:

Affinity chromatography:

Desalting:

Buffers:

HisTrap HP

Binding buffer:

Elution buffer:

GSTrap 4B:

Binding buffer:

Elution buffer:

HiPrep 26/10 Desalting:

Buffer:

HiPrep™

16/10

HiPrep™

16/10

HiPrep™

16/10

HiPrep™

16/10

Run 1 Run 2 Run 3

HisTrap HP HiPrep 26/10 Desalting

GSTrap 4B HiPrep 26/10 Desalting

GSTrap 4B HiPrep 26/10 Desalting

HisTrap HP HiPrep 26/10 Desalting

96

66

45

30

20

14

(Mr × 103)

LMW

mar

kers

Flow

thro

ugh

run

1El

utio

n ru

n 1

Flow

thro

ugh

run

2El

utio

n ru

n 2

Flow

thro

ugh

run

3El

utio

n ru

n 3

(His)6-GFP-GST

SDS-PAGE

Dua

l-tag

ged

prot

eins

Products featured: HisTrap HP, StrepTrap HP, ÄKTAxpress

Strep E. coli

Summary

188

9862

49

38

28

(Mr × 103)

1714

6

3

Dual-tagged Strep-tag II-(His)6

Mol

ecul

ar w

eigh

t mar

ker

Flow

thro

ugh

Elut

ed p

ool

Flow

thro

ugh

Elut

ed p

ool

Elut

ed p

ool

Flo

wth

roug

h

Individual HisTrap HP

HisTrap HP + StrepTrap HP

Individual StrepTrap HP

Sample: Strep

E. coli

Individual HisTrap HP purification

Column:

Binding buffer:

Elution buffer:

Flow rate:

Individual StrepTrap HP purification

Column:

Binding buffer:

Elution buffer:

Flow rate:

Two-step HisTrap HP and StrepTrap HP purification

Column:

Binding buffer:

Elution buffer:

Flow rate:

Column:

Sample:

Binding buffer:

Elution buffer:

Flow rate:

System:

SDS-PAGE

Acknowledgements:

Dual-tagged proteins

Products featured: StrepTrap HP, HisTrap HP, ÄKTAxpress, ExcelGel SDS Gradient 8–18, Multiphor II Electrophoresis System, LMW-SDS Marker Kit

Strep

Columns:

Sample: Strep E. coli

Binding buffer (StrepTrap HP):

Elution buffer (StrepTrap HP):

Binding buffer (HisTrap HP):

Elution buffer (HisTrap HP):

System:

Summary

0

100

200

300

400

500

600

700mAU

70 80 90 100 110 ml

A 280

StrepTrap HPelution

HisTrap HPelution

HisTrap HPflow through

66

30

Mr × 103

96

45

20.1

14.4

(His)6-mCherry-Strep-tag II

0

500

1000

1500

2000

mAU

70 80 90 100 110 120 130 140 ml

A 280

HisTrap HPelution

StrepTrap HPelution

Loopwash

StrepTrap HPflow through

Mr × 103

96

45

20.1

14.4

66

30(His)6-mCherry-Strep-tag II

imagination at work

Ela

nder

s i U

ppsa

la 2

008