PDF电子书：Western Blotting Handbook and Troubleshooting Guide(version 2)

dahui

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Introduction  1* Z& v) ]: e/ i# W7 J$ y
Western Blotting Overview  2-3! H" c! j: r. r7 _8 k
Step 1 – SDS-PAGE
4 F) \3 s* _2 h9 I6 `- A3 L  Thermo Scientifc Precise Protein Gels  4
+ ~! L' ]% W( ^. e6 N. d  Molecular Weight Markers  5-6
" g3 X$ c1 R1 G, X7 ~$ L) SStep 2 – Electro-Transfer
1 e( h# R1 L' V2 L9 Z$ ], _  Thermo Scientifc Pierce Fast Transfer System  7
% m  }' o1 k' s. U9 R9 T3 t  Transfer Buffers  8 * k$ ]$ a; a' K
  Filter Paper for Blotting  8 9 j; X6 Q* |; ^% v2 b, D+ \* s: X
  PVDF and Nitrocellulose Membranes  9
3 [% U; x5 u1 L; n  Thermo Scientifc Pierce Protein Stains   9  
" B3 g  h  A; b3 S0 p4 E   for Membranes   
; C( X" w2 B3 C. n7 s: W. o9 r  Antibody Extender Solution NC  10 # ~) ], E: S1 H/ N8 K. ~# }, r
  Western Blot Signal Enhancer  117 M8 j- P3 p9 H! i. V$ f* A
Step 3 – Blocking
8 ^: s: y5 N- F% g! p  Introduction  12 3 @$ v; _# w- U# w
  Blocking of Nonspecifc Binding Sites on  12
+ V3 E% k6 b* Q9 f! g   Transfer Membranes   
! O: {4 q" }  X0 \& z+ u  Blocking Buffer Optimization  12-13 % G3 `$ l' r$ O7 x, u: F# Z; X
  Blocking Buffers  13-164 v2 s9 R" l7 f
Step 4 – Formulate Wash Buffers : A% {+ T; T1 d3 w% Z2 k8 R- N
  Washing the Membrane  17 " b* k4 E$ z5 Q4 a, [/ f
  Wash Buffers  17) |# z6 q, C9 O
Step 5 – Detection Reagents % @  h- v; U" C  J. J9 @
  Validated Primary Antibodies  18-19 & f8 b* }+ e& t
  Affnity-purifed Antibodies  20
5 J3 ^( z2 z6 b! s! R1 b  Stabilized HRP Conjugates  21 1 _& z5 R. s0 U; L7 k% t5 ?
  Thermo Scientifc DyLight   22 0 J2 q0 W1 i) y7 w& C$ P! g% M* Z" I* N0 K
   Fluor Conjugates   ) m* q! `3 U  j7 w  P
  Conjugate Stabilizer Solutions  22
( e- ?; h" z6 J' m+ u  Thermo Scientifc DyLight-labeled Highly  23
% \" f5 N4 K0 P  E% q3 d( s% n* J6 v   Cross-Adsorbed Secondary Antibodies 9 t. ~9 L# J* L
  Antibody Storage and Stabilizer Solutions  24
( F, S* j: @; g4 }  Secondary Antibody Ordering Table  25-26  ; A: C0 e- }! T) n9 q
  Thermo Scientifc Clean-Blot IP  28-29 6 e$ M+ b, Y, f1 s5 }
   Detection Reagents   
, _; |7 k  @( V) |  Protein A, G, A/G and L Conjugates  30
$ Z% b8 @, \; v; n( {- B  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 8 a: C0 ~* Q( w- p" t$ ]3 F$ ?: E% ^7 _
   and Avidin Conjugates, Y& i  q( c5 y
Step 6 – Enzyme Substrates
/ q3 Q& V. t! }. f, ?0 ]Chromogenic Substrates  34-35 ! v, L" H$ n* t
Thermo Scientifc Chemiluminescent Substrates  36-43
- `' y4 ^) P* M& i  Pierce ECL Substrate  37 7 ]! \! `" Y) \9 I- |, y
  SuperSignal Chemiluminescent   38-42 ; N" L) a$ Y* o) w- v* D
   Substrates and Kits   
# g. X. Y8 }- y' O9 c  Lumi-Phos™
) a! q# P7 M" d- N# g/ O Chemiluminescent Substrate  43 5 N# [8 K1 h6 g5 [8 S& m& j3 _  K3 G
  Quick Reference Substrate Guide  43
, h1 ?$ h" K: \9 g( H4 HFast Western Blotting Kit  44
" p, {* X1 y. x" d! z5 W8 j3 SSpecialized Western Blotting Kits  45-48
% w' i" w7 U. b4 z$ p# `  Thermo SuperSignal West Pico HisProbe Kit  45 3 X  y' ?; z9 a+ a" o
  Pierce O-GLcNAc Western Blotting Detection Kit  46
% B" Y. r# L$ L- u5 z  Thermo Scientifc DyLight 549/649  46-47  * f6 N/ j$ C: C7 H3 `
   Western Blotting Kit   
- {1 \8 D/ o# X& o9 F  @; p  Thermo Scientifc DyLight 680/800 Near Infrared  47
' `; T  m7 m0 G% s* W3 E1 Z   Western Blotting Kit   
6 m- c* d' J2 G) M! f5 E4 l2 W  Thermo Scientifc Active GTPase Pull-Down  48 * L& Z4 w, f! p1 z, K. m- A- J
   and Detection Kits     Z5 z  s4 u+ h
Far-Western Blotting  49
4 y. h2 n* b) }4 f$ g% w1 J5 t8 @9 RIn-Gel Western Detection  50-51
( o+ l- x$ J+ F- v8 D: [! b0 I  Thermo Scientifc Pierce In-Gel   51
6 _1 U3 S2 l& P0 s' {8 Y1 h   Chemiluminescent Detection Kits: `* Y0 n& [" Y7 w) j
Step 7 – Film . ~  M& u1 E" g: |
  Thermo Scientifc CL-XPosure Film  52# X2 @% w2 h4 |6 ~& b
Step 8 – Stripping Buffer
" o9 Z5 u8 d% Q0 H/ ^0 `* ]Optimizing the Signal-to-Noise Ratio  53-58 & C% b4 J- n7 z$ ^4 H
  Protocol for Stripping an Immunoblot  54 % ~; i2 i  \) d/ X) k
  Thermo Scientifc Restore  55 - c' @8 }5 H, x+ C
   Western Blot Stripping Buffers
9 d3 k* Y, M# N& |( ~: N: c+ Q' v  Thermo Scientifc Restore PLUS  56 3 s0 a) s! Z: _7 D
   Western Blot Stripping Buffers ( k* w) {' o  o2 H" ]
  Thermo Scientifc Pierce   57-58 5 A7 F, s1 g. ?- |" J1 Y* V: |
   Background Eliminator
7 u3 M6 u! S" b, bTroubleshooting Guide  59-69
; e# a! ^$ l5 }  Blotting with Chemiluminescence  59
0 s% F' E, i' G9 {; u3 C  }& e7 R4 ~  Optimizing Antibody Concentration  60-63 ; S# v4 ^5 ?: x& S2 n4 A
  Problem Guide  64-67
" g: Y; n+ Y( A, ^2 r! @5 a  Full-Length Western Blotting Protocol Using   68-69 + C+ P' K& K% k/ p" f+ B3 M
   Chemiluminescent Substrates
/ k+ T# I! C* n9 a7 i0 h  O! jRecommended Reading  70$ D% d8 |/ X5 M6 [4 H2 r0 t/ x9 H

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一枝独秀

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jiandong2001

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军医

谢谢分享！

深海寂寞鱼

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白衣小彩

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sunyucheng

看看

daviddow

thanks for sharing it

mominglily

谢谢

qlwj

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