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

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  W) v4 Q9 R) z$ {% U% Q: U7 O. J7 D# D
Introduction  14 \  @' f) X% O& ]: ]% ]& w( k1 |( M
Western Blotting Overview  2-3  B( `5 B/ H% Q! E
Step 1 – SDS-PAGE
- u3 l; |- U% a' Z: ^  Thermo Scientifc Precise Protein Gels  4 - @- D+ o  S# k7 X
  Molecular Weight Markers  5-6
6 v  j. }1 d- e4 iStep 2 – Electro-Transfer
4 |% ^) ^% [% X; f2 U  Thermo Scientifc Pierce Fast Transfer System  7
5 j  i# K/ ^- _, g' A1 @0 E  Transfer Buffers  8
: y5 B: s# ]* t& x  Filter Paper for Blotting  8
4 C) ~4 @  E( W  PVDF and Nitrocellulose Membranes  9
' b" X% v0 e* a$ o- }0 F* w  Thermo Scientifc Pierce Protein Stains   9  
4 |8 X: y! n4 e7 m) M  Z+ w- Z   for Membranes   
( z; y: y- R( ^8 E  Antibody Extender Solution NC  10   b" ]% \) w6 x4 F; ?  X2 L0 [
  Western Blot Signal Enhancer  118 {. f6 J+ {( t) o7 ~5 k
Step 3 – Blocking # o& @4 C' P8 d( E( a2 z+ B4 f' k
  Introduction  12 . Y/ M& n: M5 \; E/ N
  Blocking of Nonspecifc Binding Sites on  12 ! n2 v# t! s2 n. _2 R
   Transfer Membranes   / c  p! h5 Z% B% l# C
  Blocking Buffer Optimization  12-13
; D: t9 W+ E4 _  Blocking Buffers  13-16  y* P! \. K) h
Step 4 – Formulate Wash Buffers
! v& w$ h& U$ ^" x  Washing the Membrane  17 - ]; Y0 n* ?/ d9 X$ n$ l8 @' K& k5 X
  Wash Buffers  17
- @* T  z" ?& m5 X+ AStep 5 – Detection Reagents 7 P2 ^% p& E* G$ w3 E0 {& N0 B
  Validated Primary Antibodies  18-19 $ \& i' {, j$ d. e& W3 P
  Affnity-purifed Antibodies  20 $ [# U( r7 |0 S) J9 z
  Stabilized HRP Conjugates  21 ; P: A8 p1 G' B) [/ g
  Thermo Scientifc DyLight   22 & I2 v& @7 {1 J
   Fluor Conjugates   # l. U! d3 F" x+ i  a& v
  Conjugate Stabilizer Solutions  22
1 E6 e( h) K( B0 q1 X; H" u  Thermo Scientifc DyLight-labeled Highly  23
/ a4 i: i& _: H0 ~- K; ]& I   Cross-Adsorbed Secondary Antibodies , X) X& _3 j0 r, t9 S, ]1 w; [, q
  Antibody Storage and Stabilizer Solutions  24 5 Y; O4 u0 M1 P3 g
  Secondary Antibody Ordering Table  25-26  
5 B- Q& P% V& M2 ^  Thermo Scientifc Clean-Blot IP  28-29
  o9 j$ a  s! J2 d- Z3 p   Detection Reagents   
% S0 d$ G, f$ U5 ?$ \$ A  Protein A, G, A/G and L Conjugates  30
5 y5 z8 a+ K0 i9 i  Thermo Scientifc NeutrAvidin, Streptavidin   31-33
8 W4 ^) }, k8 g" B# V8 }! V   and Avidin Conjugates
% J2 Y. P/ }$ v4 J2 Z4 ]Step 6 – Enzyme Substrates
+ M$ Q* [* u8 u; s8 RChromogenic Substrates  34-35 ; p$ |: ^7 U7 v; \& q" ~1 X9 L+ ?
Thermo Scientifc Chemiluminescent Substrates  36-43 1 E' A: M; O- ]5 Q# N
  Pierce ECL Substrate  37
+ _" F# X3 E; w) H# p  SuperSignal Chemiluminescent   38-42 * Z# j+ M# w( f
   Substrates and Kits   / k5 J5 r0 {! j
  Lumi-Phos™5 q9 y' a+ s4 r+ G# X0 w6 ~
Chemiluminescent Substrate  43
& t; G# x( `2 b  Quick Reference Substrate Guide  43
) y+ u( a5 Y  ?. F3 TFast Western Blotting Kit  44
& t9 F. W* }/ {1 u3 `, CSpecialized Western Blotting Kits  45-48
0 E+ b/ n" l! H$ m+ W  Thermo SuperSignal West Pico HisProbe Kit  45
: ?/ e+ U  t) q; y! Z; ~  Pierce O-GLcNAc Western Blotting Detection Kit  46
* Y4 V0 p1 ~) L5 w8 [  Thermo Scientifc DyLight 549/649  46-47  
( B: l# M# @( z7 ~6 e! w+ K, \% j   Western Blotting Kit   
! t2 k. D! j3 p  Thermo Scientifc DyLight 680/800 Near Infrared  47
2 D- H% V2 p2 j7 Q   Western Blotting Kit   9 t8 N. f4 A& ~% y
  Thermo Scientifc Active GTPase Pull-Down  48 ; w& j: Z5 C0 o5 C. S
   and Detection Kits   . m9 Y% \' e1 @  b! |
Far-Western Blotting  49 0 j0 d8 q6 @2 U0 y) Q3 j
In-Gel Western Detection  50-51 3 q- d/ h. R, {. I; f6 K2 D" O! |
  Thermo Scientifc Pierce In-Gel   51
) T" ]1 }# N$ I/ S7 i   Chemiluminescent Detection Kits+ ?3 \; g1 ^3 T. _/ h
Step 7 – Film
* M* M0 Y8 {/ `2 U- }7 d  Thermo Scientifc CL-XPosure Film  524 l- N7 {+ Z5 G
Step 8 – Stripping Buffer ) s  t$ x! ?" ]2 M, B! Z: {$ f
Optimizing the Signal-to-Noise Ratio  53-58 5 {. f9 [* P" Z4 X
  Protocol for Stripping an Immunoblot  54 - m" D$ h6 y8 P2 \
  Thermo Scientifc Restore  55
& |9 k* q5 C. C$ |0 N   Western Blot Stripping Buffers , s. L( n/ ?; @/ b/ e8 L
  Thermo Scientifc Restore PLUS  56 " M  m& N, l  ~5 g: j/ \
   Western Blot Stripping Buffers
& j# X6 o0 n4 N' f) K  Thermo Scientifc Pierce   57-58 4 N& h8 e9 h& x  _2 X
   Background Eliminator9 m) Q& }6 c5 L3 e7 ~
Troubleshooting Guide  59-69 $ U  K5 ^! z" J/ I  T; R
  Blotting with Chemiluminescence  59 $ t2 x2 O) w% i/ c' i
  Optimizing Antibody Concentration  60-63
/ D% r8 J: B5 F3 h. I  Problem Guide  64-67
1 w4 E* n0 {2 k* J+ {/ Y7 M  Full-Length Western Blotting Protocol Using   68-69
% y7 c% M$ Q" {5 ^" M- n7 y   Chemiluminescent Substrates & r8 V% o7 U8 e, L( ]: s
Recommended Reading  70: h3 e" [# {. {& U. K# H2 k

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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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