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

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  s% M' e8 I2 L% w! v' ?Introduction  1/ f; D6 W3 _9 z. K+ a
Western Blotting Overview  2-33 @) d; ?- L$ k& x, C  `8 C
Step 1 – SDS-PAGE
/ C4 [& P* S. I  Thermo Scientifc Precise Protein Gels  4 - k3 w( }# H0 L0 ~
  Molecular Weight Markers  5-6$ o( s- C9 t+ x% p) A/ Y. L
Step 2 – Electro-Transfer " @0 _/ T% P  e6 d! `" E
  Thermo Scientifc Pierce Fast Transfer System  7 2 |6 ~+ y6 f. B; h' t
  Transfer Buffers  8 3 y6 Y8 G/ A# I) g3 Z! D- M9 U! Y
  Filter Paper for Blotting  8 2 j0 a6 `7 t+ s( B+ e' O1 @$ h# G
  PVDF and Nitrocellulose Membranes  9
( D" f& ~& l  m7 Z2 Q- ]6 n  Thermo Scientifc Pierce Protein Stains   9  
& ]8 W/ y- E& @: j2 ?* w  U   for Membranes   ' l8 E: Q* M5 e% b- F
  Antibody Extender Solution NC  10
( i! x: u1 V* a  v' Y' y  Western Blot Signal Enhancer  11. y+ i. t% q! M5 b9 L- O
Step 3 – Blocking
5 D8 ?0 Z  Z% |7 r  Introduction  12
' |  ]7 m% f* v+ u  Blocking of Nonspecifc Binding Sites on  12
8 ]! p- }2 P( R6 l5 \   Transfer Membranes   
- ~8 V+ U. O% x+ _  Blocking Buffer Optimization  12-13
- a( S% @3 w' g  r5 ]3 R  Blocking Buffers  13-16% p0 P* n4 e6 ]6 ?
Step 4 – Formulate Wash Buffers
( B  A& X. i9 I& M, S, d, I  Washing the Membrane  17
, f0 Y8 A/ q* R9 j5 V( d0 ~2 i  Wash Buffers  175 b' b: G$ R- H& e
Step 5 – Detection Reagents 4 `3 B8 B2 Z5 p
  Validated Primary Antibodies  18-19
1 T+ f1 K# j. p7 k  Affnity-purifed Antibodies  20 % w; ?3 J. t8 Z% ~$ M5 @) u
  Stabilized HRP Conjugates  21
3 @5 A$ X; T; d* u& @, T  Thermo Scientifc DyLight   22
+ w$ u6 b) h( f' t6 m   Fluor Conjugates   
1 b1 @: q1 a. s3 J2 i$ V# f  Conjugate Stabilizer Solutions  22
* s4 {, P% A) P. V  Thermo Scientifc DyLight-labeled Highly  23 , M+ V$ Y% C) H
   Cross-Adsorbed Secondary Antibodies
/ M1 ^2 a6 D6 |  Antibody Storage and Stabilizer Solutions  24
- w. y4 S- c% L  Secondary Antibody Ordering Table  25-26  8 i, t1 x2 d2 E7 i1 Z/ u( O
  Thermo Scientifc Clean-Blot IP  28-29 % S9 h$ N1 U' K: M* D* L
   Detection Reagents   
/ p; T$ k7 j) `1 B& }; l8 S  Protein A, G, A/G and L Conjugates  30
( f6 l  t# Z9 l: r! [5 z  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 + V9 d0 P& C( X4 a9 q1 m
   and Avidin Conjugates
# d) E3 ]* M, DStep 6 – Enzyme Substrates
. \2 p1 o; O8 wChromogenic Substrates  34-35
2 ~; q1 W- g% P1 J! }* X! QThermo Scientifc Chemiluminescent Substrates  36-43
9 A5 _3 y4 `. Y+ y# }  q  Pierce ECL Substrate  37
# v: C0 D* C- c4 e  U8 ]. ^  SuperSignal Chemiluminescent   38-42 " q3 v& z+ W& k# ~) [) l
   Substrates and Kits   1 F  P$ Q, j1 m
  Lumi-Phos™
0 n. M! E; U' P+ q" J2 G9 v Chemiluminescent Substrate  43
4 P' {# v! i2 @7 F  Quick Reference Substrate Guide  43
- `" f5 w" O$ s; s$ a  nFast Western Blotting Kit  44
# x* `" n; e8 hSpecialized Western Blotting Kits  45-48
$ x  e" F7 g8 l& ^+ P& o4 s  Thermo SuperSignal West Pico HisProbe Kit  45   ?1 u% U5 y1 d6 }
  Pierce O-GLcNAc Western Blotting Detection Kit  46 ' f4 y* [' W: g) l2 O( A
  Thermo Scientifc DyLight 549/649  46-47  
3 ]0 P6 ]4 [7 o0 L) h5 M% U   Western Blotting Kit   
8 y& Q) \/ W! i/ T6 l. |3 Y) w  Thermo Scientifc DyLight 680/800 Near Infrared  47
6 x* }$ s0 t- P: B& I& P' q   Western Blotting Kit   
: _) N& }8 |8 v. y( N# _/ Y3 _8 z  Thermo Scientifc Active GTPase Pull-Down  48
' X1 p5 x2 z: S   and Detection Kits   8 e3 Y& B) U) B0 x3 Z' I+ R8 W
Far-Western Blotting  49 , z: `* V$ }: w0 Z/ N6 H- k' s  Z
In-Gel Western Detection  50-51
  R, |0 ?0 y4 R  Thermo Scientifc Pierce In-Gel   51
* E  A' s# V/ v, [, C  [   Chemiluminescent Detection Kits
2 M! A& J) S7 @4 NStep 7 – Film
* A3 t0 f+ i) e4 B4 b! \$ d  Thermo Scientifc CL-XPosure Film  52
+ b: e9 v* e' o0 XStep 8 – Stripping Buffer
% l. M5 Y* X2 C0 ^3 ?Optimizing the Signal-to-Noise Ratio  53-58
' t2 P( U% M+ [0 \+ d  Protocol for Stripping an Immunoblot  54 - n" y# ?- v' l0 F
  Thermo Scientifc Restore  55 $ N3 ?: ~. ~% U: X9 V
   Western Blot Stripping Buffers
$ D" Q% S9 o$ p* v+ X# o5 q  Thermo Scientifc Restore PLUS  56 . P, q5 A: j* l0 Y  w( g$ E5 }
   Western Blot Stripping Buffers - h# Y. W4 d& ?5 F5 f8 k; |" U
  Thermo Scientifc Pierce   57-58
( s! o- s) ]  A1 `   Background Eliminator% Q$ r; o) i) Y9 d6 S5 K, C& U0 l
Troubleshooting Guide  59-69
3 c5 b" W0 Y+ r1 E  Blotting with Chemiluminescence  59
! w2 |9 U+ e5 V  Optimizing Antibody Concentration  60-63 8 I( D7 X: q( z* c7 w
  Problem Guide  64-67
# ?3 I* p# E2 L; F2 l  Full-Length Western Blotting Protocol Using   68-69 3 {0 T% s# w: t. @& w
   Chemiluminescent Substrates 3 X4 G* N) j" x$ w5 e
Recommended Reading  709 r" k5 w/ n- {9 h5 x, Q. P) E

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一枝独秀

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jiandong2001

好久没有来这里了！谢谢分享

军医

谢谢分享！

深海寂寞鱼

好资料 学习ing

白衣小彩

学习学习

sunyucheng

看看

daviddow

thanks for sharing it

mominglily

谢谢

qlwj

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