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

dahui

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Introduction  1
8 l2 D$ M9 a/ \% ?6 oWestern Blotting Overview  2-3
: u" Q1 y! y9 }2 QStep 1 – SDS-PAGE
! N. B. E! b- d5 N% T9 r  Thermo Scientifc Precise Protein Gels  4 5 Z! K& ?6 T0 R" K0 d+ m
  Molecular Weight Markers  5-6
4 V  V2 u3 w& T; Z2 P1 q9 DStep 2 – Electro-Transfer - A9 o8 a. F! J7 k1 O
  Thermo Scientifc Pierce Fast Transfer System  7 ; Z8 ?/ O+ Q% I, d) `& m
  Transfer Buffers  8 9 f( N( P2 w& Y$ j7 q$ K3 Q! T
  Filter Paper for Blotting  8
, K% ~* Y* K2 r: x  PVDF and Nitrocellulose Membranes  9 & R# @- W; A3 ]# [) D
  Thermo Scientifc Pierce Protein Stains   9  # a" G2 P+ F9 r1 W
   for Membranes   
. x4 ~# z6 R* s+ h' p  Antibody Extender Solution NC  10 1 a, _) t. U5 f. \* h6 J" o6 W( ]
  Western Blot Signal Enhancer  11
- H4 j9 D/ k- NStep 3 – Blocking - d. A7 u( H; j0 S6 H2 t
  Introduction  12
$ m: r* ?2 P5 |& C( G  Blocking of Nonspecifc Binding Sites on  12
+ e4 s9 C# L; b) ^   Transfer Membranes   
+ O+ j2 w! P& r6 o% [  Blocking Buffer Optimization  12-13 + n9 Z+ r0 ^& J/ e6 q
  Blocking Buffers  13-16* o- Q. I# N! |  k9 P* ~( Y$ M+ |
Step 4 – Formulate Wash Buffers
" y2 k; R; f2 \/ y% L: U, G* T: v- Y  Washing the Membrane  17
3 M) s" J$ b4 _# h0 @  Wash Buffers  178 F: l! q& w5 a& e* o
Step 5 – Detection Reagents
$ Y1 q  D, F  G1 T1 X2 d0 |  Validated Primary Antibodies  18-19
9 v5 `% Z! G3 h  I0 r! t7 s  Affnity-purifed Antibodies  20 / G: F) r( p, \% b0 x
  Stabilized HRP Conjugates  21 / ?$ a' P. q& s( h+ C' W6 |6 g, y
  Thermo Scientifc DyLight   22 0 S' C2 X& c# T; Z% K/ v& ~, M) y
   Fluor Conjugates   : d: F1 G' x8 A$ |: x* y
  Conjugate Stabilizer Solutions  22
4 D. z0 }" @( i) a* v' O; @3 l. L' F  Thermo Scientifc DyLight-labeled Highly  23 + f8 A3 Z' c+ P& H
   Cross-Adsorbed Secondary Antibodies
6 K. q* s+ T+ F1 T. O/ \  Antibody Storage and Stabilizer Solutions  24
1 o# M( G6 i" f; T' Q  Secondary Antibody Ordering Table  25-26  
  y7 U1 r' ^5 F  Thermo Scientifc Clean-Blot IP  28-29 8 Q3 h" g- e" M8 r0 H. A
   Detection Reagents   + {- L/ ]3 l* E- n0 A0 z
  Protein A, G, A/G and L Conjugates  30 4 q) {3 L. D# L; e% C* Z7 ]% W
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 " w+ r8 Q; j. N$ ~* R
   and Avidin Conjugates
! U1 O! F" c% @! @: u3 @% n0 K) vStep 6 – Enzyme Substrates , F( u7 g6 z% j* ?
Chromogenic Substrates  34-35 5 N  P3 K& k8 [' `& M" y
Thermo Scientifc Chemiluminescent Substrates  36-43 5 I- K' E, `5 C* @, x
  Pierce ECL Substrate  37
4 m3 y; G5 v) G$ e: ]8 v/ U. W  SuperSignal Chemiluminescent   38-42 $ O& T6 N4 I# a
   Substrates and Kits   
* ?: @, ~0 S" D  Lumi-Phos™) ~3 ^: g' u0 z; G1 n* `2 ~
Chemiluminescent Substrate  43
2 K' S. o+ H2 i  Quick Reference Substrate Guide  43 % m# Z0 a$ N$ c
Fast Western Blotting Kit  44
* c+ F; M% ]! |8 W: Q  \6 p  dSpecialized Western Blotting Kits  45-48
$ z$ X* d" B- X, @+ ]" m- _  Thermo SuperSignal West Pico HisProbe Kit  45 % G. F4 B/ g) f6 h
  Pierce O-GLcNAc Western Blotting Detection Kit  46
4 B# F$ C! F2 i6 G# c  Thermo Scientifc DyLight 549/649  46-47  
8 R4 h3 L; \/ C) _   Western Blotting Kit   
8 Z/ H. G+ g  Q4 a( Z: L7 l  Thermo Scientifc DyLight 680/800 Near Infrared  47 & X8 g6 P4 c! f( K3 B( J
   Western Blotting Kit   
3 J8 j7 j& {) ^0 G1 s) x4 Z  M5 i  Thermo Scientifc Active GTPase Pull-Down  48
; V3 L$ f! ]& x8 [% l( a- @+ D   and Detection Kits   
* d% `/ d/ l" |; E: S" c" RFar-Western Blotting  49 5 g) S) m* \  x# ?, i+ w, N2 @0 ^2 S4 O
In-Gel Western Detection  50-51
6 C0 j2 ^/ x- ?8 h  Thermo Scientifc Pierce In-Gel   51
2 h+ ]" b$ h9 S  B8 b   Chemiluminescent Detection Kits3 O' N  a* m# p
Step 7 – Film
" e5 |/ R! c" k. S3 W9 M/ Q! W  Thermo Scientifc CL-XPosure Film  522 v# u; t3 }! i5 M7 H- C, f
Step 8 – Stripping Buffer . ~( y: N) k) {. v, v# a8 y# q' b
Optimizing the Signal-to-Noise Ratio  53-58 * K0 o% N% Y* O3 B
  Protocol for Stripping an Immunoblot  54 : w; N; ~/ i0 ], I$ u
  Thermo Scientifc Restore  55
6 R2 r- X( m1 l5 _   Western Blot Stripping Buffers 0 x/ `/ k8 S* \. W
  Thermo Scientifc Restore PLUS  56 2 I1 N0 J- G4 m+ }+ v$ ?% C+ o
   Western Blot Stripping Buffers
  `+ o/ _( w# s. B2 f  Thermo Scientifc Pierce   57-58 ! O8 a% e, q9 _' I
   Background Eliminator5 ?7 u% F: m4 \2 J: {" d- M
Troubleshooting Guide  59-69
$ C8 R, L! j" i4 r- |  Blotting with Chemiluminescence  59
1 B- |* f/ h; U; v  Optimizing Antibody Concentration  60-63 % k* f( j, b* \5 t% }( h  L8 x7 N/ Y
  Problem Guide  64-67
+ ?' l8 P, S& `! i! Z4 i  Full-Length Western Blotting Protocol Using   68-69 * z: |! d1 R7 V& q, I& H, D$ Y
   Chemiluminescent Substrates / y( X/ {  T; g3 N
Recommended Reading  70
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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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