|
- 积分
- 1343
- 威望
- 1343
- 包包
- 387
|
本帖最后由 细胞海洋 于 2011-3-12 17:58 编辑
9 _- ^% V c/ A8 `
* v$ n: j0 s- Y- o6 qIntroduction 1, Z" y7 n+ V; J" F0 Q; |
Western Blotting Overview 2-30 V; P+ E. c/ a+ Q/ F. S
Step 1 – SDS-PAGE
; t) o) Q) h. K2 b; o/ }9 L Thermo Scientifc Precise Protein Gels 4
, I) D% |1 [* j8 Q% } Molecular Weight Markers 5-6
8 a& w% [& c" t% L: x4 H8 ^ f5 wStep 2 – Electro-Transfer ; u. i1 _* _1 b
Thermo Scientifc Pierce Fast Transfer System 7 A2 X- n/ z4 I7 O
Transfer Buffers 8 ! z _+ D1 E% m7 W9 w& s
Filter Paper for Blotting 8 + S$ R# x" c) C
PVDF and Nitrocellulose Membranes 9 $ R. h6 d+ c5 ^- G% p5 u: F }
Thermo Scientifc Pierce Protein Stains 9
1 `! p: f- L/ T |" q- X$ C for Membranes
5 G9 e8 u1 p# s: ?$ N, b3 b: Z Antibody Extender Solution NC 10 ; R5 s2 r" J5 n0 ~( I% z* ? i
Western Blot Signal Enhancer 11
- Q$ A/ |4 g5 `: @) V5 aStep 3 – Blocking
7 b ]5 y) |! V6 V! t Introduction 12 / u, W' X% ?- _4 i* r: S
Blocking of Nonspecifc Binding Sites on 12 3 v6 h. o% B% v o. m
Transfer Membranes 9 s- \% B: \; T: _& R S- D
Blocking Buffer Optimization 12-13 + i& r! s# Q; m; h9 s
Blocking Buffers 13-168 A( @+ C1 ^ ^- K6 C
Step 4 – Formulate Wash Buffers
8 b* N* [2 B# v7 p { Washing the Membrane 17
' N' Z$ W( c, b. k5 b5 ?! m Wash Buffers 17
5 V+ `. D/ z; F6 EStep 5 – Detection Reagents 4 T4 m$ l8 y7 i8 R. A( c
Validated Primary Antibodies 18-19 q- K- E! I4 h. a7 H1 W! R
Affnity-purifed Antibodies 20 . Q! U( j. S; x
Stabilized HRP Conjugates 21
: y: k. e0 s; i( M. h% C1 v- ]9 W' U Thermo Scientifc DyLight 22 % ?7 \* Z1 h1 r H5 R
Fluor Conjugates
4 T D1 A# i$ X8 P* j' c5 o, C7 m Conjugate Stabilizer Solutions 22
0 Y6 @9 } ^4 T Thermo Scientifc DyLight-labeled Highly 23
6 W; X- W, n9 v- r! m& c8 ^# N Cross-Adsorbed Secondary Antibodies
- O. ~0 y. q0 P Antibody Storage and Stabilizer Solutions 24 . W1 S; [2 ?) |# k$ ]# s! y R
Secondary Antibody Ordering Table 25-26 $ M- f0 P! i6 R! ]9 \& x$ c
Thermo Scientifc Clean-Blot IP 28-29 , U u& s: \1 o, y/ s
Detection Reagents
5 X+ i6 T% W4 E Protein A, G, A/G and L Conjugates 30
. d% r1 I9 r3 E$ c$ w+ X: Y- y8 ^ Thermo Scientifc NeutrAvidin, Streptavidin 31-33 |! P( P5 K+ r% _9 t
and Avidin Conjugates( g. j, t/ m4 E1 ^0 w6 I
Step 6 – Enzyme Substrates ( V$ `3 A. E3 i- K
Chromogenic Substrates 34-35 # \- I/ d# `: G, Y
Thermo Scientifc Chemiluminescent Substrates 36-43
; Z3 U) x1 D+ f1 L Pierce ECL Substrate 37 5 q9 u( ~2 `) t5 {
SuperSignal Chemiluminescent 38-42 . |4 m4 |9 L4 A2 @4 [
Substrates and Kits 4 ?5 O" x; M2 o
Lumi-Phos™, \8 @# `2 Q4 e2 @7 G
Chemiluminescent Substrate 43
6 Y; B! Z+ M1 l e- _. Y7 L1 [ Quick Reference Substrate Guide 43 3 Y3 b' c6 G" X% L+ R
Fast Western Blotting Kit 44
0 ?4 D% s* J- U6 x- P0 n1 ySpecialized Western Blotting Kits 45-48
& b/ W% C: p$ \' t1 }' e; l% Y" ] Thermo SuperSignal West Pico HisProbe Kit 45
$ }, I3 ~) E2 g5 S6 P4 _4 {. N Pierce O-GLcNAc Western Blotting Detection Kit 46 6 i$ t# |2 q; R* r2 ]0 e0 r
Thermo Scientifc DyLight 549/649 46-47
4 S, V) g5 ~) z% R# {5 x; W Western Blotting Kit
: k$ a9 v' S# S1 C! c$ D9 b3 u4 l) @6 @ Thermo Scientifc DyLight 680/800 Near Infrared 47
/ _' d, j- i( W Western Blotting Kit
: n3 W' J6 m) h. V8 F2 Z; X Thermo Scientifc Active GTPase Pull-Down 48 % N. h' T1 k, @ r% B$ Z4 J& u
and Detection Kits
: i: p/ p- z3 M8 |, g# g# X jFar-Western Blotting 49
8 V* c. U4 `' L# {1 RIn-Gel Western Detection 50-51
% x7 b% j9 O1 Z8 [1 Y4 x: ^: J Thermo Scientifc Pierce In-Gel 51 - d' d, _5 D, j- \
Chemiluminescent Detection Kits
8 P1 ^. j0 s* u, wStep 7 – Film
3 @2 b; ^5 N8 d7 v' g2 C8 N& y; ~: k Thermo Scientifc CL-XPosure Film 52
7 u Y( x! ~0 O# jStep 8 – Stripping Buffer
# G+ f; K$ T! T7 NOptimizing the Signal-to-Noise Ratio 53-58
; x; W. I$ w8 a+ y- s( i6 a& X) T Protocol for Stripping an Immunoblot 54 " |& c! x6 H4 }+ \, C4 w$ p
Thermo Scientifc Restore 55
2 a9 z' ` n2 u5 J j5 Z9 v& a5 n Western Blot Stripping Buffers
& f3 o- z: p e1 y6 E Thermo Scientifc Restore PLUS 56 # `8 I7 a- r& ~4 ~+ S J N5 Y' T
Western Blot Stripping Buffers
3 C; V- c7 ^' f) M Thermo Scientifc Pierce 57-58
8 n5 [ X, N9 K: m- j Background Eliminator2 [6 V" E1 x# ^# `$ `) G
Troubleshooting Guide 59-69
9 v1 q5 `9 B! L+ f' { Blotting with Chemiluminescence 59
% q1 U4 @5 a; J# p! J Optimizing Antibody Concentration 60-63
M/ X2 s) a: J9 b Problem Guide 64-67
* ~3 I1 \8 \7 Z5 m( m" O& K Full-Length Western Blotting Protocol Using 68-69 ) O, E+ c- o7 A, Z: _- B, B G* O
Chemiluminescent Substrates " |" t# K. V: I" d% d6 f
Recommended Reading 70. O& s5 @1 M" u! M
% x1 ~& H2 q6 ^9 t5 d
[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
-
总评分: 威望 + 5
包包 + 10
查看全部评分
|
发表于 2011-3-12 09:56
