双向电泳实验指南

双向电泳实验指南

2-D electrophoresis for proteomics

sample preparation discussion

the first dimension: isoelectric focusing (IEF)

the second dimension: SDS-PAGE

detection of proteins in gels

detection of proteins on western blots

2-D Electrophoresis for Proteomics

image acquisition and analysis

identification and characterization of 2-D protein spots

sample solubilization and preparation methods

A Methods and Product Manual

first-dimension separation methods

second-dimension separation methods

methods for protein detection in gels

troubleshooting guide

ordering information

references and related Bio-Rad literature

双向电泳实验指南

Proteome WorksBio-Rad 50 20 50 Bio-Rad

TM

System

PCR

Rotorfor

Model 491

PAGE

Duoflow

Clonis

PROTEAN IEF

Bio-Rad

SDS PAGEBio-RadMini-Protean 3 cell 1-2 Protean II XL cell 1-2 Mini-PROTEAN 3 Dodeca cell (1-12) PROTEAN Plus Dodeca cell (1-12)

Bio-Rad

FX Pro Plus

VersaDoc

GS800

ChemiDoc XRS

PDQuest

Spot Cutter

The ProteomeWorks system is the global alliance between Bio-Rad Laboratories, Inc. (USA) and Micromass, Ltd. (UK), dedicated to furthering proteomics research.

WorkBase

双向电泳实验指南

CATELOGUE

1 1 1 1 4 4 6 8 9 9 10 pl (IPG) IEF SDS MW SDS 10 10 10 12 13 15 15 15 17 18 19 19 19 SYPRO Ruby 20 20 21 21 PVDF PVDF 22 22 23 23 24 Densitometry 25 25 26

双向电泳实验指南

CATELOGUE

28 28 29 MALDI-TOF-MS ESI-LC-MS-MS MALDI-MS-MS 29 30

31 31 31 34 35 37 37 38 41 Precast Gels IPG IPG 41 42 42 43 44 R-250 Bio-Safe SYPRO Ruby Bio-Rad Merril Silver Stain plus Stain G-250 44 44 44 45 45 46

47 51

Bio-Rad" 2-D Electrophoresis for proteomics Manual" (Bulletin 2651) 021-64260808

A Methods and Product

http://www.77cn.com.cn

双向电泳实验指南

Proteomics Proteome

(

) 1.1

2-D PAGE 1,800 2000) ( Choe and Lee

(Cutler et al. 1999, Fegatella et al. 1999, Gorg et al. 2000)

1

双向电泳实验指南

!2 !9

isoelectric point pI pI pH IEF pH IPG !3 !21! SDS !(MW)

4 5 6

7

Bio-Rad !PDQuestTM

双向电泳实验指南

!!!! !

!!!

Bio-Rad !(ExQuestTM spot cutter) 7

ExQuest 96 PDQuest ProteinChip SEL-/ SELDI Surface Enhanced Laser Desorption/Ionization SELDI ProteOn 36

双向电泳实验指南

!!!! ! !!!

5-8M 2M Rabilloud 1998

Efufshfout pI SDS octylglucoside CHAPS CHAPSO 1-2% Rabilloud 1999 SB3-10 ASB-14 Chevallet et al. 1998 !4% Hermann et al. 2000

pH 5–8 strip

!P,!

!40mM ǘ !IPG! !0.2% w/v carrier ampholytes, pH 5–8/8–10 (2:1), 80,000 V-hr

2.1 ψ

!2.1! pH 17cm pH5-8 Readystrip IPG

SDS PAGE 10-24 with PDA crosslinkerIPG! pH5-8 A : pH3-10 carrier ampholytes

B : pH5-8 carrier ampholytes : pH8-10 carrier ampholytes 2 :1

!A! pH3-10! , !B! !pH5-8! !pH8-10! !2:1

双向电泳实验指南

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