PDF电子书：蛋白质组学最新版Introduction to Proteomics [Liebler[1].Humana.2002] - 经典PDF电子书下载区干细胞之家



免疫细胞治疗专区	欢迎关注干细胞微信公众号

12 3 4 5 6 7 8 9 10 ... 25 下一页

返回列表

查看: 700701\|回复: 245	go [生物学相关学科类] PDF电子书：蛋白质组学最新版Introduction to Proteomics [Liebler[1].Humana.2002] [复制链接]

dahui

高级会员

Rank: 4

积分: 1343
威望: 1343
包包: 387

金话筒优秀会员热心会员小小研究员积极份子帅哥研究员

楼主

发表于 2010-6-27 21:10 |只看该作者 |倒序浏览 |打印

本帖最后由细胞海洋于 2010-6-28 11:09 编辑 & L* |% @$ H8 S' E2 H

Proteomics in Practice2 M1 K( {7 b; n* e. m8 ]
A Guide to Successful Experimental Design4 A4 |/ d9 @( h) M7 _% D5 S

Abbreviations, Symbols, Units XV
Introduction 1# q8 ?6 s. J7 L& B
1 History 1$ v" J9 Q$ Z3 R+ N8 S  {) m
2 Critical Points 8+ s. J/ m" S9 U6 p; O
2.1 Challenges of the Protein Samples 8
2.1 Challenges of the Analysis Systems 11' ~$ J% R7 |* _: t! i- l
3 Proteomics Strategies 125 x) o1 {; U# p9 U$ Y  D( V' ^4 X: [
3.1 Proteome Mapping 12
3.2 Differential Analysis 12
3.3 Time Point Experiments 13+ }" u0 i, n- T% u
3.4 Verification of Targets or Biomarkers 13% w; h1 R% U+ f) U0 G8 d
3.5 Integration of Results into Biological Context 13
3.6 Systems Biology 132 v0 `) w% X0 g( o
4 Concept of Experimental Planning 14
4.1 Biological Replicates 14% U$ N: c4 u- N
4.2 Pooling of Samples: Yes or No? 14$ R+ G( L- r2 M% I( M, O! ^
4.3 Pre-fractionation of Samples: Yes or No? 141 z" ^9 c  S) ?* u% C6 c
4.4 Which is the Best Workflow to Start With? 15
Part I: Proteomics Technology
1 Electrophoretic Techniques 19
1.1 The Principle of Electrophoresis and Some Methodological/ K; R/ t/ D, |7 f- _
Background 19
1.1.1 Free Flow Electrophoretic Methods 20
1.1.2 Gels for Electrophoretic Techniques 21+ h( i% G4 ]4 C
1.1.3 Electroendosmosis Effects 21
1.2 Polyacrylamide Gel Electrophoresis 22: i3 s9 B5 p% p# A

1.2.1 The Polyacrylamide Gel 22
1.2.2 SDS Polyacrylamide Gel Electrophoresis 270 ^2 |! N& I/ C
1.2.3 Blue Native Electrophoresis 32
1.2.4 Cationic Detergent Electrophoresis 34, D: W8 t* u: s5 h: F
1.3 Blotting 35
1.3.1 Electrophoretic Transfer 36
1.3.2 Protein Detection on the Membrane 36
1.4 Isoelectric Focusing 386 t" v  ]% l8 I; |9 a: l, X
1.4.1 Theoretical Background 39% ]9 {% a  k  y, q/ t9 Q( q
1.4.2 Preparation of IEF Gels 44/ U9 R* g( s6 Y: T- j
1.4.3 Isoelectric Focusing in Proteomics 451 w8 l7 Y5 {, O9 P' w9 ^9 N4 l/ F' j2 Y
1.5 Two-dimensional Electrophoresis 53
1.5.1 Sample Preparation 53
1.5.2 Pre-labeling of Proteins for Difference Gel Electrophoresis 68
1.5.3 First Dimension: Isoelectric Focusing in IPG Strips 77
1.5.4 Second Dimension: SDS Electrophoresis 1003 m- q, d6 ]+ d. R
1.5.5 Detection of Protein Spots 119+ k; g" n6 i6 w, |" ]- u
1.6 Image Analysis 1251 j& a, `; t/ W3 T: V! N
1.6.1 Image Acquisition 125% S1 N9 C' l3 X7 W5 ?
1.6.2 Image Analysis and Evaluation 129! a& t, z9 Z; O) u6 q- k
1.6.3 Use of 2-D Electrophoresis Data 137
1.7 Spot Handling 137
1.7.1 Spot Picking 139/ x1 i9 @& Z1 K# h2 F6 y, v
1.7.2 Protein Cleavage 141/ U6 y+ H1 k% Q- _2 I% u4 q
Liquid Chromatography Techniques 151
2.1 Basic Principles of Important Liquid Chromatography: `4 ]6 }) y2 w% t, w
Techniques 151
2.1.1 Ion Exchange Chromatography 153$ `/ W+ U1 X4 }5 D$ W, E5 v0 v) d" |
2.1.2 Reversed Phase Chromatography 162
2.1.3 Affinity Chromatography 167) W: g: U: [& V4 U
2.1.4 Gel Filtration 172
2.2 Strategic Approach and General Applicability 174
2.3 Liquid Chromatography Techniques and Applications in Proteome
Analysis 176
2.3.1 Peptide Separation 176
2.3.2 2DLC Peptide Separation 179
2.3.3 Affinity Chromatography and LC-MS/MS 187
2.3.4 Protein Pre-fractionation 1895 E( z  e$ Y. t
2.4 Practical Considerations and Application of LC-based Protein: i+ T& N. c+ m, {( L5 g
Pre-fractionation 194
2.4.1 Sample Extraction and Preparation 1964 o+ V/ X- I% x7 ]
2.4.2 Experimental Setup 197
2.4.3 Ion Exchange Chromatography and! _4 M+ H( p; e$ V+ [# w$ i
Protein Pre-fractionation 198
Contents
2.4.4 Reversed Phase Chromatography and
Protein Pre-fractionation 205
2.4.5 Fraction Size and Number of Fractions 210
2.5 Critical Review and Outlook 211
3 Mass Spectrometry 215* }) M) }& B" x& c8 B
3.1 Ionization 218
3.1.1 Matrix Assisted Laser Desorption Ionization 218
3.1.2 Electrospray Ionization 222
3.2 Ion Separation 2251 S* F- f8 h5 G" l
3.2.1 Time-of-Flight Analyzer 225; d- i' M3 w. J5 s5 E
3.2.2 Triple Quadrupole Analyzer 227
3.2.3 Quadrupole Ion Trap 228
3.2.4 Quadrupole Time-of-Flight 230' B: |- m+ t! F% U
3.2.5 Hybrid Triple Quadrupole Linear Ion Trap 231
3.2.6 TOF/TOF Analyzer 231
3.2.7 Fourier Transform Ion Cyclotron 232
3.2.8 Orbitrap 233
3.3 Generating MS Data for Protein Identification 233
3.3.1 Peptide Mass Fingerprint 2346 B1 j- L- y0 `- w( D8 r
3.3.2 Peptide Mass Fingerprint Combined With Composition
Information 237: s1 B. x/ r8 N+ f( \8 c
3.3.3 Peptide Mass Fingerprint Combined With Partial Sequence8 y9 M& K/ Q% q! W4 q3 V& _0 b: i
Information 238
3.3.4 Tandem Mass Spectrometry 242
3.4 Protein Characterization 2582 v9 ~# r& o8 M- N) ^- ^
3.4.1 Phosphorylation Analysis 259
3.4.2 Affinity Chromatography 260
3.4.3 Chemical Derivatization 261
3.4.4 Glycosylation 263. p; g/ L; L7 C9 D9 u9 F9 c
3.5 Protein Quantification Using Mass Spectrometry 264& h3 k! ^1 d$ p: D1 I, V
3.5.1 Stable Isotope Labeling Approaches 2646 g, Y3 G# h+ K( E
3.5.2 Isotope-coded Affinity Tags 265
3.5.3 Stable Isotope Labeling with Amino Acids in Cell Culture 2663 |4 p( @6 A5 L2 W9 R
3.5.4 AQUA 267
3.5.5 iTRAQ 2676 j2 \! |. t( T3 E% Z( Z
3.5.6 Non-labeling Software Approaches 268
3.6 MS Strategies 271
3.6.1 Bottom up Approach 271: U( p3 F) j2 [
3.6.2 Top down Approach 272/ s8 Z2 p7 k* K0 B* A% c  {
4 Functional Proteomics: Studies of Protein–Protein Interactions 273
4.1 Non-immunological Methods 2739 q8 [( `" i7 r& b/ n; u
4.1.1 Separation of Intact Multi-protein Complexes 273: h# G+ z6 o) Q
4.1.2 Probing with Interaction Partners 2734 Z7 s  U. y( j4 I
4.1.3 Surface Plasmon Resonance 274( \9 O4 R( V: M  N# _+ Q' _! Z+ S
4.2 Antibody-based Techniques 275
4.2.1 Western Blotting and Dot Blots 275
4.2.2 Protein Microarrays 276
Part II: Practical Manual of Proteome Analysis 279
Equipment, Consumables, Reagents 281; f& l. m+ Q" l9 y0 y: v: p/ ^
Step 1: Sample Preparation 287
Step 2: Fluorescence Difference Gel Electrophoresis 299" ?5 X( B5 V5 {
Step 3: Isoelectric Focusing 309
Step 4: SDS Polyacrylamide Gel Electrophoresis 323
Step 5: Scanning of Gels Containing Pre-labeled Proteins 357
Step 6: Staining of Gels 3614 X0 T/ \1 n1 v& i  S
Step 7: Image Analysis and Evaluation of DIGE Gels 373
Step 8: Spot Excision 3830 j# e' d' B. Y" `- P. T. ~: ]
Step 9: Sample Destaining 387) t0 R7 b" u$ V0 S$ o
Step 10: Protein Digestion 389. N, ^0 G# n- w, K! p
Step 11: Microscale Desalting and Concentrating of Sample 393
Step 12: Chemical Derivatization of the Peptide Digest 3976 B8 K1 o, Y9 q6 J# c8 T: y3 q
Step 13: MS Analysis 399
Step 14: Calibration of MALDI-ToF MS 403
Step 15: Preparing for a Database Search 407
Part III: Trouble Shooting 411$ j4 u, C; i. p4 x: K4 F
1 Two-dimensional Electrophoresis 413
1.1 Sample Preparation 413' z" t' \. w9 E/ c
1.2 Isoelectric focusing in IGPG strips 414
1.3 SDS PAGE 416
1.4 Staining 4176 s& Q, f" J: [* Z! K& q3 ?& c, M
1.5 DIGE Fluorescence Labeling 418
1.6 Results in 2-D Electrophoresis 421
2 Mass Spectrometry 429
6 l: F: J5 N' b; ]4 T: J
[hide][/hide]

附件: 你需要登录才可以下载或查看附件。没有帐号？注册