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

dahui

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3 t4 [! m4 M8 M0 T$ W% r) Q1 [Introduction  15 \$ P: r8 u* N- F
Western Blotting Overview  2-3
7 t) \9 k. P0 H$ e. qStep 1 – SDS-PAGE $ ]9 k* O8 }: ]; O' ~
  Thermo Scientifc Precise Protein Gels  4 5 {# D! O$ M* ]1 i0 |3 g
  Molecular Weight Markers  5-6$ K& `* A; n6 Y$ a% r8 S3 w
Step 2 – Electro-Transfer
% ?# m, s- N( ~/ o% H  Thermo Scientifc Pierce Fast Transfer System  7
, s+ {2 i1 d% u; v3 Z& j. u  Transfer Buffers  8
$ U# I- Q, P  \- x: j& _  Filter Paper for Blotting  8
* W3 w. Z1 r8 z1 Y; n* `  t  PVDF and Nitrocellulose Membranes  9
+ z' F$ K# T+ p) c  Thermo Scientifc Pierce Protein Stains   9  
' v) O' B2 @7 Q   for Membranes   1 M3 w3 s* i! ]% z3 Q) h( l0 T
  Antibody Extender Solution NC  10
; Y- x4 x4 H  I* R: f$ J  Western Blot Signal Enhancer  112 X& e8 B1 g' s) U' N+ G
Step 3 – Blocking
4 w# {- Z9 K, Z$ ]* x* s5 _' A7 N  Introduction  12
$ N9 @/ U/ n1 t1 S) d7 t( A: Z' O: {  Blocking of Nonspecifc Binding Sites on  12
% _  o9 S0 J! b# @/ A; p   Transfer Membranes   
: J0 L( q0 h8 r  D/ D' F1 B( e  Blocking Buffer Optimization  12-13 . L* q  Z3 F7 w3 J% @
  Blocking Buffers  13-16
4 ?% D" g7 }: Q2 |Step 4 – Formulate Wash Buffers
: P/ x3 \* r8 G; N0 M* g  Washing the Membrane  17
/ F- |9 }% b* y. Q* o/ d+ k  Wash Buffers  176 j9 d2 {6 _9 W3 {* F
Step 5 – Detection Reagents 0 l! V4 a' L% ]& l7 g, |/ u
  Validated Primary Antibodies  18-19 ; x9 ?* }0 d- x1 ?. u
  Affnity-purifed Antibodies  20 8 \' K$ _5 _3 c- ]
  Stabilized HRP Conjugates  21 8 T4 p; T3 L! w* Y# L& [
  Thermo Scientifc DyLight   22
0 \  }% `1 P! T; H9 l5 K   Fluor Conjugates   
1 G; u* t0 x: _  Conjugate Stabilizer Solutions  22
8 H- r! B$ Z) B" @, i7 C7 S  Thermo Scientifc DyLight-labeled Highly  23 & M' ?. }. Q9 v3 C+ |" ~; C' M. W
   Cross-Adsorbed Secondary Antibodies
( N& v  J& c& D: U# Z8 j  Antibody Storage and Stabilizer Solutions  24
1 D3 ~; v) ?1 m% m  Secondary Antibody Ordering Table  25-26  ; S$ h0 y& ^. e
  Thermo Scientifc Clean-Blot IP  28-29
: |( ~# r) P2 |- `8 D+ Q" e   Detection Reagents   % X9 a. A# i, D% Y! M7 _
  Protein A, G, A/G and L Conjugates  30 & {$ ~) l/ F7 M' o' B1 I& V
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 ) k1 e6 Z/ t# \& g6 C6 ?/ Q! H
   and Avidin Conjugates
8 l# s, G; c: E1 v6 O; |Step 6 – Enzyme Substrates
3 H+ I6 n. W4 g: M+ ?) nChromogenic Substrates  34-35
) M% N  U6 s2 @! L. p( C3 qThermo Scientifc Chemiluminescent Substrates  36-43 , X. \$ _/ n5 S# j1 _
  Pierce ECL Substrate  37 / M# P: ]( {0 E$ n8 f7 K
  SuperSignal Chemiluminescent   38-42   C1 Y# W# U: b, M
   Substrates and Kits   
2 p  s) W2 J3 q& E  Lumi-Phos™
' ^) m) s2 n% _ Chemiluminescent Substrate  43
4 v1 F# x) H& `# y/ H  Quick Reference Substrate Guide  43 7 w8 ^. B. e* {$ g0 W
Fast Western Blotting Kit  44 $ R5 q, V' K) Q. i+ R" h: t* R
Specialized Western Blotting Kits  45-48
2 s5 P* G2 T( y1 ]  Thermo SuperSignal West Pico HisProbe Kit  45
. z: M. l. _; z9 p% x  Pierce O-GLcNAc Western Blotting Detection Kit  46 7 r8 G5 S$ }* K$ B
  Thermo Scientifc DyLight 549/649  46-47  $ I9 i5 F) Z$ A, `0 T
   Western Blotting Kit   1 Q- \9 _$ k. y' z: K4 }/ @
  Thermo Scientifc DyLight 680/800 Near Infrared  47 # ^8 t" E5 M4 B& R: I
   Western Blotting Kit   
# v6 L1 ]& J* x' u  Thermo Scientifc Active GTPase Pull-Down  48
+ Z* t3 P7 |% I$ ]" l! Q   and Detection Kits   
4 V& D" ^; T+ ~( ~& N. w/ z8 aFar-Western Blotting  49 8 Z/ R! l( c5 F4 E& F5 }
In-Gel Western Detection  50-51
0 N. Z) \+ C# x, r& d4 Y  Thermo Scientifc Pierce In-Gel   51 ' |3 h; a0 q" G& X4 V
   Chemiluminescent Detection Kits4 Q  S& w) `" o) M% L0 d: k. z
Step 7 – Film
5 Y, s: l+ A, r: W5 L, O8 W9 W5 q  Thermo Scientifc CL-XPosure Film  529 N4 h7 ]0 n0 ~
Step 8 – Stripping Buffer
# ?8 P3 u& R! n$ H7 }5 M' Q4 HOptimizing the Signal-to-Noise Ratio  53-58 4 \7 i! q$ a" i. s# D
  Protocol for Stripping an Immunoblot  54
& h7 m2 o; k) e* R  Thermo Scientifc Restore  55
7 v6 n) r1 }& ^$ }8 k4 a- r& i7 i   Western Blot Stripping Buffers / M, B( Q0 P2 k. C* I' o$ j
  Thermo Scientifc Restore PLUS  56
* h8 j, `% k; ~8 W- i   Western Blot Stripping Buffers
. I- ^3 t; `9 r2 M) A  Thermo Scientifc Pierce   57-58
! D6 l( O( c4 l% E$ [   Background Eliminator
6 u0 V6 B1 W& S3 g: M* KTroubleshooting Guide  59-69
# @8 |, E$ Q* v  Blotting with Chemiluminescence  59 9 ^% i5 D  [( H0 s$ w, Q! ~, b
  Optimizing Antibody Concentration  60-63 , `, P3 x* |  t, P/ b
  Problem Guide  64-67 ) {* ]/ b' U$ \1 a( l
  Full-Length Western Blotting Protocol Using   68-69 % A6 m/ u# I3 d7 |3 k5 \
   Chemiluminescent Substrates
! F- l# a5 i% F, xRecommended Reading  70; b# Z1 o3 y0 S" ?2 c
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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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