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

dahui

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7 ^( a1 V* g5 r" Z) j+ Q! N$ PIntroduction  1
, ]$ H' S8 C2 L- S& ?& l" \Western Blotting Overview  2-3
2 z* [+ F+ d$ i- i9 E! QStep 1 – SDS-PAGE
; R& F8 A2 u/ v0 ]  Thermo Scientifc Precise Protein Gels  4
$ K  C* S  F6 B; S) f  Molecular Weight Markers  5-6( D: t/ \3 F( ~' c  |5 U1 [
Step 2 – Electro-Transfer / L" P/ f) s: t  o% E/ X8 e0 k
  Thermo Scientifc Pierce Fast Transfer System  7 & S) B) ]: @0 X
  Transfer Buffers  8 . q; C9 V. ]% G: S
  Filter Paper for Blotting  8 4 M7 M  a' A9 k1 ]( l
  PVDF and Nitrocellulose Membranes  9
5 x. V6 W+ E2 G8 o5 E9 I7 Y$ t  Thermo Scientifc Pierce Protein Stains   9  
/ D- o8 i  r3 E1 T& G   for Membranes   
6 M8 B% t2 A% _4 y  l& Z  Antibody Extender Solution NC  10
: v7 _3 E4 g- J- @  ]0 y2 Q  Western Blot Signal Enhancer  112 s( {8 |8 R6 M3 w" t4 @1 h$ D4 g
Step 3 – Blocking * k0 X" C6 p( D+ p
  Introduction  12
4 S& L/ {6 @8 @6 ^$ e  Blocking of Nonspecifc Binding Sites on  12 9 W8 [/ F; V7 \& L8 g% Q
   Transfer Membranes   
8 i3 p# J' {1 c0 ]  Blocking Buffer Optimization  12-13 / p: R  s7 B3 o) P3 ?( P
  Blocking Buffers  13-16
% m8 X1 g' `2 g: KStep 4 – Formulate Wash Buffers
6 d4 ]( n  a5 R+ ~  t  a9 }: G  Washing the Membrane  17
/ Y8 E; x$ P2 W" _, w% o4 w  Wash Buffers  17, L; c7 x5 G! J
Step 5 – Detection Reagents & W+ e# `4 k) ]$ l6 \
  Validated Primary Antibodies  18-19
( }7 V/ l- l1 u7 O$ ~  Affnity-purifed Antibodies  20 ; Q6 H9 e/ N% r1 m. P7 n
  Stabilized HRP Conjugates  21 2 \" S1 E! R; G2 W% [
  Thermo Scientifc DyLight   22
- u* u4 Y+ |  v& ]- i   Fluor Conjugates   8 Q& F# F+ y- n+ d9 k. d
  Conjugate Stabilizer Solutions  22 / T' \/ P5 m# B# S# G# i2 L
  Thermo Scientifc DyLight-labeled Highly  23
( U$ O! C* F; f9 a% n   Cross-Adsorbed Secondary Antibodies , b8 Q7 V) N8 m7 }4 u2 l) _/ S+ R$ p
  Antibody Storage and Stabilizer Solutions  24 9 y4 p! b; [# ]1 U* [- |
  Secondary Antibody Ordering Table  25-26  
+ Y# z8 _. S" @% J; `$ \1 C  Thermo Scientifc Clean-Blot IP  28-29 + w8 {4 ~2 d; b) S7 ~) R  }) M
   Detection Reagents   
0 Z& y2 N0 ^2 b% w  Protein A, G, A/G and L Conjugates  30
: }+ ?& h8 ]" `! L' S' d  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 : `* |8 x, Q! c" c+ [6 ?
   and Avidin Conjugates
/ N' L2 ], }4 Q$ _, l+ IStep 6 – Enzyme Substrates ; D, J% o1 m! r/ ]3 S; X
Chromogenic Substrates  34-35
+ ~% g0 [) v) D; w# `% MThermo Scientifc Chemiluminescent Substrates  36-43 1 h; F* I$ x2 |! ]2 w1 R& N1 Q
  Pierce ECL Substrate  37 ( l8 H) O; W8 O( w  Y& M1 P
  SuperSignal Chemiluminescent   38-42
( h4 h8 \, M% S+ o+ g+ B   Substrates and Kits   
2 k! ^! Q4 A4 F* P1 N  Lumi-Phos™
5 I+ f  f% a) q  j Chemiluminescent Substrate  43
& _1 t  c% z" A' ~; C# ]  Quick Reference Substrate Guide  43 . Z, `4 x& L) i* o* z
Fast Western Blotting Kit  44 3 F3 g. I6 D8 q1 k" e4 r% a) W. y
Specialized Western Blotting Kits  45-48
9 c6 o$ g' {3 M9 y' A+ H# N0 ], E. b, n  Thermo SuperSignal West Pico HisProbe Kit  45
2 b6 P( @- d8 l; Y4 J  Pierce O-GLcNAc Western Blotting Detection Kit  46 ; t6 B/ X6 ]# l7 ]% C
  Thermo Scientifc DyLight 549/649  46-47  
3 J/ t% B4 A- w  |3 S' M6 k' y   Western Blotting Kit   3 H+ q5 p( o5 N. S: B
  Thermo Scientifc DyLight 680/800 Near Infrared  47 . y- H( I1 j* }. Q) Y8 D& C* `
   Western Blotting Kit   
: y: D# y8 {6 C  Thermo Scientifc Active GTPase Pull-Down  48 7 k8 e# }! k* \# e+ o
   and Detection Kits   , R# l9 t3 e1 b
Far-Western Blotting  49 4 b$ u7 G  l  a+ Y- ^( b  T+ M& n
In-Gel Western Detection  50-51
0 l4 Y! w2 R  Q, ]/ u, \  Thermo Scientifc Pierce In-Gel   51
4 W; n" T6 |$ H   Chemiluminescent Detection Kits
( @  f& B8 C7 K4 oStep 7 – Film
1 G$ H3 Y! x/ N: v# Q3 ?  Thermo Scientifc CL-XPosure Film  52% c6 a* x9 N2 y1 f) c. P
Step 8 – Stripping Buffer 0 W/ Y4 u& `) R* p0 A9 I& t
Optimizing the Signal-to-Noise Ratio  53-58
% h4 u. @- P- O  Protocol for Stripping an Immunoblot  54 * F, m( S% V- |1 g5 U! {
  Thermo Scientifc Restore  55
- A! \' P0 {* u: R. _& c   Western Blot Stripping Buffers 1 a, n0 r4 B, W2 ^
  Thermo Scientifc Restore PLUS  56 ) T. u0 v7 b5 L
   Western Blot Stripping Buffers ; c8 L# f. m; N* B1 x, ]0 `
  Thermo Scientifc Pierce   57-58
7 R7 B: F% e: I" F* @   Background Eliminator) G& U+ t. C  ^% G
Troubleshooting Guide  59-69
2 e, d1 N' @4 o( M8 H4 s* H  Blotting with Chemiluminescence  59 + b" \* A3 F) ?/ X
  Optimizing Antibody Concentration  60-63 % r1 z, D* z/ c* b+ J- i
  Problem Guide  64-67 ! x6 |# Z8 u! }5 Z
  Full-Length Western Blotting Protocol Using   68-69   ]/ O, W3 m# u9 j1 H
   Chemiluminescent Substrates   q4 F( u# p" u& y/ D, J' p
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

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mominglily

谢谢

qlwj

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