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

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Introduction  1
& w/ ?) s  C; b9 ~6 LWestern Blotting Overview  2-3
& [, c1 j: ^& o1 {Step 1 – SDS-PAGE
7 a1 N% F' ~8 `7 i3 `  Thermo Scientifc Precise Protein Gels  4
) y: t# a0 Z( N6 x  Q  Molecular Weight Markers  5-6) o8 k7 J  b9 `% Z1 R: @
Step 2 – Electro-Transfer
/ _; q& Q" Q5 n+ q( t9 ]* X  Thermo Scientifc Pierce Fast Transfer System  7 0 ~7 i* D$ T, _# }- r
  Transfer Buffers  8 ' j' t2 K& t1 k! j- r
  Filter Paper for Blotting  8
& u* |' u$ r  }+ |3 a7 O+ f  PVDF and Nitrocellulose Membranes  9 # ^+ v5 X2 r, m! {0 E2 n
  Thermo Scientifc Pierce Protein Stains   9  
6 o, r  o* K8 w   for Membranes   
" L- e! P7 @% G" c8 T  Antibody Extender Solution NC  10
/ x5 |' F% ]+ C) m; ~  Western Blot Signal Enhancer  11
* i# f* d% c" F; x" [% ^Step 3 – Blocking
5 q* P6 ^1 U7 ~6 _+ b" D  Introduction  12
! H* ~* I" \/ Y) F  Blocking of Nonspecifc Binding Sites on  12 . I- f+ A' x" j9 S8 E
   Transfer Membranes   8 v" Z, [5 T) L4 u& p6 |. A9 X9 A
  Blocking Buffer Optimization  12-13
4 m( Y/ g' F& P7 E' l( E  Blocking Buffers  13-164 D; v6 m! N! ^2 B* o
Step 4 – Formulate Wash Buffers : T3 q+ U3 A$ d0 K
  Washing the Membrane  17
: t* L. }9 Q+ Y2 Q. n  Wash Buffers  17
# S4 K7 h' L; I. T8 EStep 5 – Detection Reagents , u$ N2 ~1 {6 I' N- h6 l' C6 R+ m% K
  Validated Primary Antibodies  18-19 0 d/ P8 X5 S1 x0 ^2 ]$ O
  Affnity-purifed Antibodies  20 : |3 ]% `- g* C7 o4 O2 ~# e8 ?# u
  Stabilized HRP Conjugates  21
) W2 k; Y! `1 h/ \& s  Thermo Scientifc DyLight   22 : k( x% K4 }1 f( n
   Fluor Conjugates   
7 R* V8 d9 N, B  Conjugate Stabilizer Solutions  22
& i. [! G$ N/ A. Z8 R  Thermo Scientifc DyLight-labeled Highly  23
! Y0 I3 A6 f6 p! F1 a   Cross-Adsorbed Secondary Antibodies ) ]0 U# |7 S* `; \5 X
  Antibody Storage and Stabilizer Solutions  24 8 i# V: I7 S  N- K) b0 A
  Secondary Antibody Ordering Table  25-26  ' i- ?. s! s! U
  Thermo Scientifc Clean-Blot IP  28-29 2 m; E4 v) [2 _9 o
   Detection Reagents   3 p, C" |- Y9 h
  Protein A, G, A/G and L Conjugates  30 4 q' u  }( q, n& H# f, t
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 3 H  P1 Y/ Z# S9 Z& r  h! h; Z
   and Avidin Conjugates8 z3 |! E9 Z5 z' ?
Step 6 – Enzyme Substrates 1 h4 A5 D' @5 s' C) c. e  @* v
Chromogenic Substrates  34-35
, r& U4 f  `& a) ~) S: kThermo Scientifc Chemiluminescent Substrates  36-43
9 k" a3 d" e' F# i% _: R+ g0 X  Pierce ECL Substrate  37 0 u$ f8 u; m/ _: \9 c
  SuperSignal Chemiluminescent   38-42
( l) o. u4 }2 M; Q( @% o' t& g/ d   Substrates and Kits   
& z1 m$ x/ h: d  Lumi-Phos™
7 o2 _; h8 K2 r0 [- D$ Q Chemiluminescent Substrate  43 * _9 n! A" @: q9 W
  Quick Reference Substrate Guide  43
: x/ I& Z  O9 R* z. i* l; v+ dFast Western Blotting Kit  44
/ e2 H7 J3 _8 h+ QSpecialized Western Blotting Kits  45-48
+ r' O6 _- ^% [9 A0 D+ r  Thermo SuperSignal West Pico HisProbe Kit  45
1 r! R6 t+ d# _! _% u  ^3 C6 [; S$ S  Pierce O-GLcNAc Western Blotting Detection Kit  46
  ?- w; _* M2 K0 U- n  Thermo Scientifc DyLight 549/649  46-47  
0 Z5 p# |/ t& ?! P   Western Blotting Kit   : {5 T! N7 Y- ^, h7 K, w) a- z
  Thermo Scientifc DyLight 680/800 Near Infrared  47 7 G/ n7 d( u6 U$ H& [
   Western Blotting Kit   7 _) V: j! v/ d
  Thermo Scientifc Active GTPase Pull-Down  48 0 q8 Y( R4 ?& s; |- E" N
   and Detection Kits   * H0 u1 H( ~/ V7 @) K8 N! u$ W' W
Far-Western Blotting  49
7 |9 s( i+ N4 ^  ]4 E" bIn-Gel Western Detection  50-51 7 b/ t% g5 Z3 E  Y; k8 W9 C
  Thermo Scientifc Pierce In-Gel   51 # L/ l! X4 }4 a/ F5 g; s
   Chemiluminescent Detection Kits
$ p4 w$ p% ]+ ?Step 7 – Film 4 H) X. `4 J: m( ~
  Thermo Scientifc CL-XPosure Film  52
1 N* j! G& O+ c9 Q! U1 \Step 8 – Stripping Buffer 9 b' y7 B4 N# M
Optimizing the Signal-to-Noise Ratio  53-58 : Q( }6 V8 X7 l% \0 i. _
  Protocol for Stripping an Immunoblot  54
; t) }- u5 q" T( {5 \  Thermo Scientifc Restore  55 4 W7 ^2 K1 t9 a& a2 ^$ `9 _) J
   Western Blot Stripping Buffers 1 p" [7 ~8 g) J0 F; s
  Thermo Scientifc Restore PLUS  56
3 ?; M& g3 i" p$ h7 s   Western Blot Stripping Buffers
6 g1 E& z$ _) c) d+ e4 ^, O  Thermo Scientifc Pierce   57-58 - |3 _5 d* l4 k2 N3 {3 z8 }
   Background Eliminator
0 g# C4 q) P, Z# |" zTroubleshooting Guide  59-69
2 U0 L7 |& z# X# D: E& T) T  Blotting with Chemiluminescence  59
+ C2 o2 T0 ]$ F+ g  Optimizing Antibody Concentration  60-63 3 f6 z+ ?$ u! G
  Problem Guide  64-67
8 G5 D8 E$ t# c  Full-Length Western Blotting Protocol Using   68-69
# v' F6 a/ C- P( i; q" P1 a   Chemiluminescent Substrates
1 B- O5 R4 v, Y4 j' L7 n2 U! GRecommended Reading  70, E9 G% W9 k' E0 j+ ~

2 A1 K* Y" K) d* H; {! F: X9 b6 C4 r[hide][/hide]

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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