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

dahui

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Introduction  1
- Q$ B5 G" q& [# \1 gWestern Blotting Overview  2-3
# i- c7 a% c$ F  x/ }7 v. }Step 1 – SDS-PAGE ' M& e. G: {- ~- ]9 J
  Thermo Scientifc Precise Protein Gels  4 + _8 x8 O+ `! P4 s1 o; x
  Molecular Weight Markers  5-6
: J. s/ O$ g1 o, _) I. GStep 2 – Electro-Transfer
3 n9 e3 a) ?8 d  Thermo Scientifc Pierce Fast Transfer System  7
4 T8 Y4 i, i& j* ]  Transfer Buffers  8
% [! p+ U2 h* |; A! W! U  Filter Paper for Blotting  8 1 a8 m# X' Y4 g7 s
  PVDF and Nitrocellulose Membranes  9 + s0 N% L; J0 N+ M# k& I
  Thermo Scientifc Pierce Protein Stains   9  
* }$ _& J$ R, y4 i* C: q8 a   for Membranes   + k* H1 w( N& r
  Antibody Extender Solution NC  10
0 _# @7 S3 \, U+ l) r, Y  Western Blot Signal Enhancer  11' [! J0 c, Y' F; o  E5 K: ?2 }" J
Step 3 – Blocking
4 Y7 o1 H/ _! J: H  Introduction  12 ! H0 m, J1 u5 P) g* c! ]9 t* k. v* w
  Blocking of Nonspecifc Binding Sites on  12 , N+ ^- |3 F- D1 a& D* |
   Transfer Membranes   
7 n6 P6 v) b" [  I5 }2 j  Blocking Buffer Optimization  12-13
1 t; x6 r( \& e5 N8 t  Blocking Buffers  13-16
1 j% A: J# o* @6 S  EStep 4 – Formulate Wash Buffers ) X9 u7 F; P9 I$ d9 w7 b
  Washing the Membrane  17 8 W: h7 Z0 V0 _- {$ W; B8 X+ q. Z6 S
  Wash Buffers  17" d0 v& p7 ~+ L. k* B
Step 5 – Detection Reagents ' i0 d6 }6 a6 i# u( y$ \; K4 {, H
  Validated Primary Antibodies  18-19 , z- n* _1 d: d) V
  Affnity-purifed Antibodies  20   ?& {: a' ^" V! U- L8 v% F/ k# c
  Stabilized HRP Conjugates  21
$ Y4 l2 V& c$ `5 P  Thermo Scientifc DyLight   22 ; z) I6 {/ w; t% o' E9 ]3 a
   Fluor Conjugates   0 f; _* h# w  Y- Q. g0 y3 h
  Conjugate Stabilizer Solutions  22 ' w* c7 e- l0 D+ O6 B# h
  Thermo Scientifc DyLight-labeled Highly  23 3 I& F. O2 V2 B% J6 o. p! {
   Cross-Adsorbed Secondary Antibodies
. X( A* P) F, h& Z( R+ g  Antibody Storage and Stabilizer Solutions  24 # B% g3 b1 J& n4 E# W( x# D
  Secondary Antibody Ordering Table  25-26  
/ h3 P$ |: q; s- R0 O  Thermo Scientifc Clean-Blot IP  28-29
# K7 Z/ t, @* I9 |# J   Detection Reagents   ! n8 T  L( b$ |! o" f- ^
  Protein A, G, A/G and L Conjugates  30
/ N% _+ q0 J% C; F0 {/ k& M  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 0 x* P+ {1 \2 v  ?
   and Avidin Conjugates7 r" a' I8 u8 s# ?) o, r3 W2 F1 V
Step 6 – Enzyme Substrates $ u: U$ o; K' L0 o9 i: \; ]
Chromogenic Substrates  34-35 6 M4 O5 b$ U$ O1 T7 K3 _/ O
Thermo Scientifc Chemiluminescent Substrates  36-43 $ l/ u/ ~% `; I/ u/ u
  Pierce ECL Substrate  37 $ B+ r2 y) a; S! f0 M0 X( N1 E% H
  SuperSignal Chemiluminescent   38-42 7 B" t1 F& z1 Q) Z' N" ^
   Substrates and Kits   # b' i" U: R' M% B) t# f; H
  Lumi-Phos™
8 }7 a% d9 O2 G! a Chemiluminescent Substrate  43
4 A5 ~4 ^; C2 m4 `) K  Quick Reference Substrate Guide  43 : Y6 A" N! t* p* G2 x! g% I  u
Fast Western Blotting Kit  44   k/ G. {+ _7 v) a" \
Specialized Western Blotting Kits  45-48
5 u# G4 P! K- Z: k4 H5 n, P  Thermo SuperSignal West Pico HisProbe Kit  45
# P) p' U# N/ i  \+ O  Pierce O-GLcNAc Western Blotting Detection Kit  46
2 L$ L, U1 J4 X3 W6 |1 e  Thermo Scientifc DyLight 549/649  46-47  
2 c& P5 f3 k9 E) k" m1 K/ g   Western Blotting Kit   
. Z% Y0 ?) P( y7 `6 c$ g  Thermo Scientifc DyLight 680/800 Near Infrared  47
) v2 T1 |0 k2 ?% H0 }; {   Western Blotting Kit   
/ @; U% e0 @' }1 j" B  w  Thermo Scientifc Active GTPase Pull-Down  48 7 i" [' e. I) H% g! c
   and Detection Kits   9 H: O0 p! h4 n9 o8 F6 s
Far-Western Blotting  49
* K' o: G5 m( e; o. `In-Gel Western Detection  50-51
1 Z. g2 w3 e. u$ @# Y* c  Thermo Scientifc Pierce In-Gel   51 2 B* l3 v" p6 V/ z& t
   Chemiluminescent Detection Kits/ Y( n9 ?6 T# {9 X
Step 7 – Film : f1 ]1 J9 r8 y$ M- I+ V
  Thermo Scientifc CL-XPosure Film  52) W# s  X9 ~/ U9 t, J0 b8 t
Step 8 – Stripping Buffer
6 L+ Q9 P7 e4 x! i2 A9 UOptimizing the Signal-to-Noise Ratio  53-58 2 m( t. E" G% z8 b  v  H
  Protocol for Stripping an Immunoblot  54 3 |$ D' E$ V% |$ i: K( G' M
  Thermo Scientifc Restore  55 & R' H' R1 `# s% s
   Western Blot Stripping Buffers
( _& K- o7 c$ ^9 x0 c7 W4 i  Thermo Scientifc Restore PLUS  56
1 X$ \! K! B# @   Western Blot Stripping Buffers 8 ~& D2 \. O8 @. h6 j
  Thermo Scientifc Pierce   57-58 4 I0 V) \$ W5 M, `$ K
   Background Eliminator/ f4 _/ L0 U  F' E) p
Troubleshooting Guide  59-69
( d  g4 h. ?  u* Z  Blotting with Chemiluminescence  59
  a7 D: O# p2 h, x  Optimizing Antibody Concentration  60-63 ; R6 r  d( @) B3 m/ G
  Problem Guide  64-67 8 a9 B& g- e  i4 {+ i* P5 w* a8 l
  Full-Length Western Blotting Protocol Using   68-69 ) r" M( O! \" \6 M
   Chemiluminescent Substrates
) \. l( o+ S$ {Recommended Reading  70
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一枝独秀

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jiandong2001

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军医

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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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