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

dahui

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$ J+ W" ~7 U/ j3 WIntroduction  15 f1 p% j! d' B9 @1 z
Western Blotting Overview  2-39 U2 y& K/ B# H1 X- x9 u
Step 1 – SDS-PAGE ) F9 c7 }% ]0 J, j, J* z
  Thermo Scientifc Precise Protein Gels  4 5 T# K) z- U+ b8 ], t0 A$ [
  Molecular Weight Markers  5-6, G% q% B( f7 M
Step 2 – Electro-Transfer
! L' _, |9 N! ~  Thermo Scientifc Pierce Fast Transfer System  7 4 Y. F* c8 M  s
  Transfer Buffers  8 4 M* l' S2 @4 d5 S0 N* D0 q+ X  _
  Filter Paper for Blotting  8 , ?, p- {9 k' N3 ^4 r) V& Q! E
  PVDF and Nitrocellulose Membranes  9
6 `: Q( [7 }" [  Thermo Scientifc Pierce Protein Stains   9  
& H% o, S6 v/ c8 ~4 R; ]   for Membranes   ( G9 H0 {9 p0 C/ K; k1 x$ ]
  Antibody Extender Solution NC  10
4 w4 U5 C& _" q& X  Western Blot Signal Enhancer  11
; j3 v3 U  z- DStep 3 – Blocking . _* @' M- b. `) Y
  Introduction  12
% O- M' l( A6 K3 i: U  Blocking of Nonspecifc Binding Sites on  12 . I3 B- e, E/ U# d5 {: F# }5 D
   Transfer Membranes   $ b- {! a' D1 T7 _- I2 b! }
  Blocking Buffer Optimization  12-13
! w/ w( R* [. d; w  Blocking Buffers  13-16
  y% \, h5 s: l* v5 ]5 ]: kStep 4 – Formulate Wash Buffers ( V" u- H4 o' I
  Washing the Membrane  17 ! M% q% g8 M8 y5 R) j2 E
  Wash Buffers  17
: \1 q9 m. G5 q/ _; KStep 5 – Detection Reagents ; S; t  {2 ?" z- `( h
  Validated Primary Antibodies  18-19
6 K  A" q# F# F- ]" J0 x0 F  Affnity-purifed Antibodies  20
: I0 p: r$ \0 Q  Stabilized HRP Conjugates  21 1 q% b/ c' A/ Y6 C. ^9 n+ \9 ~
  Thermo Scientifc DyLight   22 * T2 i- V: s/ A7 O, y0 z
   Fluor Conjugates   / a: z3 O7 G7 ]" f! A* b
  Conjugate Stabilizer Solutions  22
- H% e8 Y) O+ B3 `: ?  Thermo Scientifc DyLight-labeled Highly  23
$ x# k9 @$ O, ]4 l. s   Cross-Adsorbed Secondary Antibodies
$ ]: ^* O. m7 J  Antibody Storage and Stabilizer Solutions  24 7 g: `3 l  K& o. k* u
  Secondary Antibody Ordering Table  25-26  6 W, |, L9 G4 x" {( P  S6 N# f
  Thermo Scientifc Clean-Blot IP  28-29
& |+ |* M( u; G( {6 }9 P   Detection Reagents   
1 j" R* g( t, `0 x  Protein A, G, A/G and L Conjugates  30
, o+ ]0 ]% o* c& O9 j# C* |$ s  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 + y9 d: k2 X  E; h) ?! n8 W2 V: X
   and Avidin Conjugates
: O' B( I9 [1 g" B- k7 w- J: d1 N! pStep 6 – Enzyme Substrates , ]1 \) B9 b. K; c! P
Chromogenic Substrates  34-35
1 }! ?9 Y, H% Q9 ?  N4 aThermo Scientifc Chemiluminescent Substrates  36-43 0 i, R# u( w. Y0 {: k* i7 S
  Pierce ECL Substrate  37 ; }+ v/ [9 x. ~3 o- @2 a
  SuperSignal Chemiluminescent   38-42 ! z- J1 r) Q, q+ R' T8 V4 W1 U
   Substrates and Kits   1 }/ n, D" N' i" s
  Lumi-Phos™) E. G8 Q+ D. a6 t: ?# y; \
Chemiluminescent Substrate  43
  R- ~# Z, u$ t, J& i0 R2 Z  Quick Reference Substrate Guide  43
7 `: k( s/ p) O! }Fast Western Blotting Kit  44
" u% s2 F; Q. n9 p+ bSpecialized Western Blotting Kits  45-48 0 C( b& u( s: x# k6 \
  Thermo SuperSignal West Pico HisProbe Kit  45 2 x5 A$ c4 j) U1 I" {
  Pierce O-GLcNAc Western Blotting Detection Kit  46 * V6 J2 X' J; G# D% S0 R. ^! z
  Thermo Scientifc DyLight 549/649  46-47  
0 }8 h5 P5 G1 ?! ]+ r1 z   Western Blotting Kit   
- @) W3 _3 A- A9 e9 v$ n. \: O- t  Thermo Scientifc DyLight 680/800 Near Infrared  47   |" u! [" @' ^4 F/ |' d/ A( s- D0 k
   Western Blotting Kit   
" }. B, z5 J0 ^. a; r  Thermo Scientifc Active GTPase Pull-Down  48
% B* S- n7 T4 n1 ~! @' w  D6 Q   and Detection Kits   & |& `! b( v! U
Far-Western Blotting  49 & Z2 a# X0 P/ ?
In-Gel Western Detection  50-51 - C9 }% z; R4 j6 u; K. T4 f! n
  Thermo Scientifc Pierce In-Gel   51 8 Z+ v  ?; p  Q' N8 C1 i
   Chemiluminescent Detection Kits4 T' ~5 {) e. C, b- p! v) v. v
Step 7 – Film : A! c" w4 i# n" C% A8 L" v- }6 [2 E. e
  Thermo Scientifc CL-XPosure Film  52
+ g1 n# ]9 n/ o7 T8 }/ ]6 DStep 8 – Stripping Buffer . X/ i+ w0 c- Q) ]9 q2 m! g9 N
Optimizing the Signal-to-Noise Ratio  53-58
% w* Z; @$ M8 i% M+ ~  Protocol for Stripping an Immunoblot  54 ) t; Y' k- `4 @  X
  Thermo Scientifc Restore  55
/ @( H# y9 b) R' Z( Y$ y) V   Western Blot Stripping Buffers $ B8 c& q2 T& A
  Thermo Scientifc Restore PLUS  56
& t& \# B# I- q   Western Blot Stripping Buffers 7 a; V0 X9 R3 F$ g/ K
  Thermo Scientifc Pierce   57-58 9 r. }0 {# x( T/ h
   Background Eliminator
( A* _* M7 z1 O# c7 v) T1 m) w& v4 tTroubleshooting Guide  59-69
7 E4 ~6 m& ?, X2 E- D1 C- A% q1 I  Blotting with Chemiluminescence  59 8 _& Q7 _/ X( d- G
  Optimizing Antibody Concentration  60-63 " L( R0 t; W+ v& R
  Problem Guide  64-67
) y  `) o: _! K+ e  Full-Length Western Blotting Protocol Using   68-69
3 W) N/ N/ E; [/ v7 B   Chemiluminescent Substrates $ Z! a/ Z9 ]4 I6 n/ G1 o
Recommended Reading  70' a$ ^3 W" U- j# I6 {1 c: p9 b

  U1 C, L- s* c( t+ W1 ?$ F! g[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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