Gel Running Buffer.docx

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Gel Running Buffer.docx

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Gel Running Buffer.docx

GelRunningBuffer

AlthoughTris-glycineSDS-PAGEgelsarethemostcommonmethodforseparatingproteinsforanalysis,anygelseparationtechniquecanbeused.Othertechniquesforproteinseparationinclude:

isoelectricfocusing,SDS-PAGEusingbuffersotherthanTris-glycine,urea-polyacrylamidgels,tris-borateandnativelow-saltpolyacrylamidegels.Allthebuffersarepreparedwithhighqualityreagents.Electrophoresisbuffersaresoldasconcentrate.Pleasedilutewithdeionizedwatertomake1X. PleaseNote:

Thecompositionof1Xisprovidedunlessspecifiedotherwise.

M0195-MESSDSRunningBuffer.500ml,sterilesolution.500ml

Description:

3.2gMES（2-[N-Morpholino]ethanesulfonicacid）monohydrate,121.2gTrisBase,20.0gSodiumDodecylSulfate,7.631gEDTA.

Product

CatalogNo.

Quantity

Price

IEFAnodebuffer（50X）,pH2.4

Phosphoricacid

7mM

BP-210

500ml

1000ml

$15

$25

IEFCathodebuffer（10X）pH10.1

Arginine（freebase

20mM

Lysine（freebase）

20mM

BP-211

250ml

500ml

$35

$65

IEFCathodebuffer（10X）pH3-7

Lysine（freebase）

20mM

BP-214

250ml

500ml

$35

$65

MES-SDSrunningbuffer（20X）,pH7.3

MES

50mM

Trisbase

50mM

SDS

0.1%

EDTA

1mM

BP-177

500ml

1000ml

$42

$80

MOPS-SDSrunningbuffer（20X）,pH7.7

MOPS

50mM

Trisbase

50mM

SDS

0.1%

EDTA

1mM

BP-178

500ml

1000ml

$42

$80

Tris-Acetate-SDSrunningbuffer（10X）,pH8.24

Trisbase

50mM

Tricine

50mM

SDS

3.5mM

BP-179

500ml

1000ml

$30

$55

Tris-Glycine-SDSrunningbuffer（10X）,pH8.3

Tris-base

25mM

Glycine

192mM

SDS

0.1%

BP-150

1000ml

2000ml

4000ml

$21

$37

$70

Tris-Glycine-nativerunningbuffer（10X）,pH8.5

Tris-base

25mM

Glycine

192mM

BP-160

1000ml

2000ml

4000ml

$21

$37

$70

Tris-Tricine-SDSrunningbuffer（10X）,pH8.3,（Cathodebuffer）

Trisbase

100mM

Tricine

100mM

SDS

0.1%

BP-170

500ml

1000ml

$37

$70

Tris-Tricine-SDSrunningbuffer（10X）,pH8.9,（Anodebuffer）

Tris-HCl

200mM

BP-172

500ml

1000ml

$21

$37

Tris-Tricine-SDSbuffer（10X）kit

ThekitcontainsoneeachofBP-170andBP-172

BP-175

500ml

1000ml

$45

$85

Product

CatalogNo.

Quantity

Price

TBErunningbuffer（10X）,pH8.3

Trisbase

89mM

Boricacid

89mM

EDTA

2mM

BP-180

1000ml

2000ml

4000ml

$21

$37

$70

ZymogramRenaturationbuffer（10X）

TritonX1002.7%（w/v）inDIwater

BP-217

500ml

1000ml

$15

$25

ZymogramDevelopingbuffer（10X）

Tris-HCl（pH,7.5）

50mM

NaCl

200mM

Cacl2

5mM

Brij35

0.02%

BP-218

500ml

1000ml

$21

$37

BlotTransferBuffer

Transferofproteinsfromthegelontothemembraneisachievedinthreeways:

simplediffusion,vacuum-assistedsolventflow,andelectrophoreticelution.Electrophoreticelutionisthemethodofchoicebecauseofitsease,speedandcompletenessoftransfer.Electrophoreticelutioncanbeachievedeitherbywetorbysemi-drytransfer.

PleaseNote:

Thecompositionof1Xisprovidedunlessspecifiedotherwise.

Product

CatalogNo.

Quantity

Price

TransferorElectroblottingbuffer（10X）,pH8.3

Tris-base

25mM

Glycine

192mM

Justaddmethanolaccordingtoyourprotocol

BP-190

1000ml

2000ml

4000ml

$21

$37

$70

Tris-Capsbuffer（10X）

Anodebuffer,pH9.6 Cathodebuffer,pH9.5

Tris

60mM

Tris

60mM

Caps

40mM

Caps

40mM

Methanol（add）

15%

SDS

0.1%

BP-192

500ml

1000ml

$25

$45LuminolandOxidizingSolutionsforwesternblotting（HRPBased）JustmixLuminolandOxidizingsolution,incubatethemembranefor1min.andexposethex-rayfilmfordifferentperiodsoftime.WB-100250mleach500mleach$50$85

LuminolandOxidizingSolutionsforwesternblotting（HRPBased）

JustmixLuminolandOxidizingsolution,incubatethemembranefor1min.andexposethex-rayfilmfordifferentperiodsoftime.

WB-100

250mleach

500mleach

$50

$85

BlotBlockingBuffer

Beforetheblotcanbeprocessedforantigendetection,itisveryimportanttoblockthenon-specificbindingsitefortheantibodies.BBPprovidesfollowingreagentsthatareusedtoblockthenon-specificsitesonthemembrane.

PleaseNote:

Thecompositionof1Xisprovidedunlessspecifiedotherwise.

Product

CatalogNo.

Quantity

Price

BSA-blockingbufferinPBS

3%BSAinPBS

IBB-100

200ml

500ml

$30

$55

BSA-blockingbufferinTBS

3%BSAinTBS

IBB-110

200ml

500ml

$30

$55

Casein-blockingbufferinPBS

3%CaseininPBS

IBB-120

500ml

1000ml

$30

$55

Casein-blockingbufferinPBS（withazide）

3%Caseinand0.02%SodiumazideinPBS

IBB-121

500ml

1000ml

$30

$55

Product

CatalogNo.

Quantity

Price

Casein-blockingbufferinTBS

3%CaseininTBS

IBB-130

500ml

1000ml

$30

$55

Casein-blockingbufferinTBS（withazide）

3%Caseinand0.02%SodiumazideinTBS

IBB-131

500ml

1000ml

$30

$55

Gelatin-blockingbufferinPBS

1%GelatininPBS

IBB-134

500ml

1000ml

$30

$55

Gelatin-blockingbufferinPBS（withazide）

1%Gelatinand0.02%SodiumazideinPBS

IBB-135

500ml

1000ml

$30

$55

Gelatin-blockingbufferinTBS

1%GelatininTBS

IBB-136

500ml

1000ml

$30

$55

Gelatin-blockingbufferinTBS（withazide）

1%Gelatinand0.02%SodiumazideinTBS

IBB-137

500ml

1000ml

$30

$55

BlottoinPBS

5%nonfatdrymilk,0.02%SodiumazideinPBS

IBB-140

250ml

500ml

$25

$40

BlottoinTBS

5%nonfatdrymilk,0.02%SodiumazideinTBS

IBB-150

250ml

500ml

$25

$40

Blotto/TweeninTBS

5%nonfatdrymilk,0.1%Tween-20,0.02%SodiumazideinTBS

IBB-160

500ml

1000ml

$25

$40

PBS/Tween-20（10X）

0.1%Tween-20inPBS

IBB-170

500ml

1000ml

$25

$40

PBS/Tween-20（10X）

0.05%Tween-20inPBS

IBB-171

500ml

1000ml

$25

$40

TBS/Tween-20（10X）

0.1%Tween-20inTBS

IBB-180

500ml

1000ml

$25

$40

TBS/Tween-20（10X）

0.05%Tween-20inTBS

IBB-181

500ml

1000ml

$25

$40

Gelatin10%

IBB-184

100ml

200ml

$35

$60

Gelatin/TweeninPBS

1%Gelatin,0.05%Tween-20,0.02%SodiumazideinPBS

IBB-185

500ml

1000ml

$40

$70

Gelatin/TweeninTBS

1%Gelatin,0.05%Tween-20,0.02%SodiumazideinTBS

IBB-186

500ml

1000ml

$40

$70

HorseSerum

10%Horseserum,0.02%sodiumazideinPBS

IBB-190

100ml

200ml

$35

$60

BlotWashingBuffer

PleaseNote:

Thecompositionof1Xisprovidedunlessspecifiedotherwise.

Product

CatalogNo.

Quantity

Price

Phosphate-bufferedsaline （PBS,10X,pH7.4）

Phosphatebuffer

10mM

KCL

2.7mM

NaCl

137mM

BM-220

1000ml

2000ml

4000ml

$21

$37

$70

PBS-Tween,（10X,pH7.4）

Phosphatebuffer

10mM

NaCl

150mM

Tween-20

0.05%

BM-221

250ml

500ml

$21

$37

Phosphate-bufferedsaline （PBS,10X,pH7.4）

Forwesternblotwashing

SodiumPhosphatebuffer

10mM

NaCl

150mM

BM-222

1000ml

2000ml

4000ml

$21

$37

$70

Tris-bufferedsaline （TBS,10X）

Tris

25mM

KCL

2.7mM

NaCl

137mM

BM-300

1000ml

2000ml

4000ml

$21

$37

$70

TBS-Tween-20,（10X.pH7.4）

Tris

50mM

NaCl

150mM

Tween-20

0.05%

IBB-181

250ml

500ml

$21

$37

Tris-bufferedsaline （TBS,10X,pH7.4）

Forwesternblotwashing

Tris

50mM

NaCl

150mM

BM-301

1000ml

2000ml

4000ml

$21

$37

$70

Tris-bufferedsaline （10X,highsalt,pH7.4）

Forwesternblotwashing

Tris

50mM

NaCl

500mM

BM-301A

1000ml

2000ml

4000ml

$25

$45

$80

Tris-bufferedsaline （TBS,10X,pH8.0）

Tris

50mM

NaCl

150mM

BM-301B

1000ml

2000ml

4000ml

$21

$37

$70

BlotStrippingBuffer

Product

CatalogNo.

Quantity

Price

Strippingbuffer,（4X）,pH6.8

Tris-HCl

250mM

SDS

Justaddmercaptoethnol

BP-96

250ml

500ml

$25

$45

Strippingbuffer2,（4X）,pH2.5

Glycine

0.2M

BP-97

250ml

500ml

$25

$45

MOLECULARBIOLOGY:

WORKINGWITHPROTEINS

PROTEINDETECTION:

GELELECTROPHORESIS

RUNNINGNOVEXNuPageGELS

RunningNovexNuPageGels

Contributor:

TheLaboratoryofSteveHahnattheFredHutchinsonCancerResearchCenter

Overview

ThesearesimplifiedinstructionsforuseofNovexprecastNuPagegels.TheXCEllII™minicellapparatusisrequiredforthisprotocol.FormoreinformationonNovexgelsandproductsseetheirwebpage

Procedure

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