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

dahui

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Introduction  1( p3 q  c8 E$ J6 d
Western Blotting Overview  2-3; s: L/ B( j" R7 `, y
Step 1 – SDS-PAGE / i; i* u! m! r" `, S1 A
  Thermo Scientifc Precise Protein Gels  4
/ A: i' w7 J3 f, {0 r  Molecular Weight Markers  5-66 ~* i# F! U) m  N& _" @! R& G
Step 2 – Electro-Transfer + R! ?+ L. Q1 J
  Thermo Scientifc Pierce Fast Transfer System  7
; S  j; X3 p9 x2 m7 x  Transfer Buffers  8
) c" r/ B+ Y1 z* M2 T  Filter Paper for Blotting  8 ' V7 k" X/ \* U4 M( C, ]
  PVDF and Nitrocellulose Membranes  9 $ C2 d+ J' O' m! D9 V$ V
  Thermo Scientifc Pierce Protein Stains   9  * a4 z' k, a+ v* ?# v0 A6 O
   for Membranes   , i# ^1 ?4 V7 h! P5 W0 E8 h
  Antibody Extender Solution NC  10 - p, H$ d& {8 ^) r0 B! J7 c5 Z5 E1 g2 R
  Western Blot Signal Enhancer  11
! {( y- m, O  S5 f0 PStep 3 – Blocking 2 B3 P* _9 Y5 m; q( ?" d9 T7 `1 N/ F
  Introduction  12
1 R' N( c) p) `3 i  Blocking of Nonspecifc Binding Sites on  12
' V' @* v' z" @   Transfer Membranes   9 ]2 ]8 a0 b7 }6 L" h
  Blocking Buffer Optimization  12-13
8 M8 P$ p+ E" U" T  Blocking Buffers  13-167 o" d$ K0 P4 w7 |0 M' C
Step 4 – Formulate Wash Buffers 3 ]+ w3 @1 R- w# P. l3 ]
  Washing the Membrane  17 6 E/ G; R& @1 G' `  K& S
  Wash Buffers  173 C$ W1 t$ R0 c
Step 5 – Detection Reagents ; ?/ ]* s8 g  D+ g! p* g
  Validated Primary Antibodies  18-19 9 m2 j) A) l- ?* z
  Affnity-purifed Antibodies  20
9 W- ?9 _4 j( H. s+ ]' n  Stabilized HRP Conjugates  21 ) k0 n- z! p2 q: D8 r
  Thermo Scientifc DyLight   22
3 @9 I, Q+ n+ n" r   Fluor Conjugates   3 I5 j# m& O9 r& X1 p1 h
  Conjugate Stabilizer Solutions  22 0 M7 Y. U: K, H1 _
  Thermo Scientifc DyLight-labeled Highly  23 2 y, G- r2 U0 Q: S
   Cross-Adsorbed Secondary Antibodies
+ u, x! p  `% k7 {& j  U' M  Antibody Storage and Stabilizer Solutions  24
: N5 w9 _7 n" V( r  Secondary Antibody Ordering Table  25-26  / g3 T0 s9 p" A3 e& n( s4 y
  Thermo Scientifc Clean-Blot IP  28-29
* E; B" N6 H2 o3 m   Detection Reagents   0 c! S% E" e# s/ E
  Protein A, G, A/G and L Conjugates  30 ) \/ t! j; ?4 _/ {; ]) [6 ^
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 ) p, @+ l+ w+ C9 ~. {# z
   and Avidin Conjugates) A# t3 y& {& k( a
Step 6 – Enzyme Substrates 2 P% A$ b) z5 s* j8 i
Chromogenic Substrates  34-35
6 {$ z  W7 B- r' rThermo Scientifc Chemiluminescent Substrates  36-43 ' E0 e: O: t6 Z/ g5 d0 M5 o
  Pierce ECL Substrate  37
" c7 O4 S% i$ ^7 f% X$ R  SuperSignal Chemiluminescent   38-42 . y  L; [0 S8 C
   Substrates and Kits   
# z; s- s- c( T: Q  Lumi-Phos™
% |) i. c* W! B' A9 v# r Chemiluminescent Substrate  43 3 l0 P) ]1 f* J% I9 Y
  Quick Reference Substrate Guide  43 + w1 u. K4 K3 q% _- N: f; y( R  p
Fast Western Blotting Kit  44 # h* N  }! `. M; I% V
Specialized Western Blotting Kits  45-48 : U1 B( O% S) `/ S% G
  Thermo SuperSignal West Pico HisProbe Kit  45
. t2 [1 x" N' b' r7 `  Pierce O-GLcNAc Western Blotting Detection Kit  46
5 H2 B7 ]8 Y; }* K  Thermo Scientifc DyLight 549/649  46-47  
$ e& A2 e! V7 G8 R$ k   Western Blotting Kit   
8 I5 G* X2 Y& F% a" j* y* Y  Thermo Scientifc DyLight 680/800 Near Infrared  47
- K$ ~% P' m9 g2 c. H8 E   Western Blotting Kit   & d& A) M, w  l" F% r0 J
  Thermo Scientifc Active GTPase Pull-Down  48 2 r( {" X" u3 _8 M
   and Detection Kits   ( R0 e; e1 ^3 O4 M1 G: A
Far-Western Blotting  49   N# \# \' \: [" U) P
In-Gel Western Detection  50-51
8 O8 l6 h" y- h! J7 b; `  Thermo Scientifc Pierce In-Gel   51   v7 U) U6 L8 d' m$ t5 P
   Chemiluminescent Detection Kits& G; w, x& K# f5 D; Y  s' s: J
Step 7 – Film " i2 D' Y6 t7 P3 R
  Thermo Scientifc CL-XPosure Film  52
* M' R0 W# I$ EStep 8 – Stripping Buffer
3 {# j: m/ q; a9 W* @Optimizing the Signal-to-Noise Ratio  53-58 : n5 v2 Q) N5 P3 u3 O* O6 a
  Protocol for Stripping an Immunoblot  54
$ H+ d' @3 h4 x* G. J  Thermo Scientifc Restore  55
4 Y' Q8 t! s8 y6 {; l+ [) h4 f   Western Blot Stripping Buffers + w1 R. q! T1 z& q
  Thermo Scientifc Restore PLUS  56
3 Y  K& m2 Z# Z) P- ?2 v   Western Blot Stripping Buffers 6 s4 y2 R- W7 @
  Thermo Scientifc Pierce   57-58 + [% i2 g  `- u7 d0 u
   Background Eliminator, l2 Q; v- D& L! Z
Troubleshooting Guide  59-69 ' i  \& s: {) N7 D" K1 W9 q: U3 ?
  Blotting with Chemiluminescence  59 8 \  V+ a  d6 L' `" J* E7 ?/ h
  Optimizing Antibody Concentration  60-63 7 a- T8 ?8 m# w6 a& Z
  Problem Guide  64-67 ) x0 }* t7 O! t6 D8 a' z$ ~
  Full-Length Western Blotting Protocol Using   68-69
5 R6 T  H% w: p: P2 r6 S. F   Chemiluminescent Substrates
  F  o# a0 {( F% u7 P( ^% ARecommended Reading  70
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一枝独秀

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jiandong2001

好久没有来这里了！谢谢分享

军医

谢谢分享！

深海寂寞鱼

好资料 学习ing

白衣小彩

学习学习

sunyucheng

看看

daviddow

thanks for sharing it

mominglily

谢谢

qlwj

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