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

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; V* Q5 h$ R* j, f# lIntroduction  1
& _! a3 w) q: D, |+ A* v1 p; B$ IWestern Blotting Overview  2-37 u( g2 I9 ^( u* B4 ]) w
Step 1 – SDS-PAGE : v, Y* N6 N% D* U' R- x
  Thermo Scientifc Precise Protein Gels  4 & V& ~* Y+ J: v8 j- ^
  Molecular Weight Markers  5-61 _) m+ Z4 Q9 h. H9 s
Step 2 – Electro-Transfer
3 ]( a) K2 V6 T  Thermo Scientifc Pierce Fast Transfer System  7
6 ^1 j! N! u/ F3 w! v% p  Transfer Buffers  8 ( O! v9 d3 |! y: ~" i5 ^
  Filter Paper for Blotting  8
; p! p$ r" P; Z0 T1 j  PVDF and Nitrocellulose Membranes  9
& m7 W5 t8 q  h6 ^% A# m' ]  Thermo Scientifc Pierce Protein Stains   9  " v% @6 B7 [1 m4 j9 H' Z) m
   for Membranes   
$ f! {* |& \+ R& k6 L1 t8 B3 D  Antibody Extender Solution NC  10 9 D$ c. o+ M8 y6 X2 W5 {
  Western Blot Signal Enhancer  11
) O9 C1 ^9 p# k% iStep 3 – Blocking 8 x, a7 l2 e5 w8 t* d' `
  Introduction  12
& Q( ?4 W  y; L2 h  Blocking of Nonspecifc Binding Sites on  12
6 m3 e- ~8 l; k- L) z' `6 ]- s   Transfer Membranes   
$ s# V) R8 F# r7 u4 A1 r5 ~  Blocking Buffer Optimization  12-13
0 |7 i8 ]5 y2 ?/ z3 y1 T' ~  Blocking Buffers  13-16
/ _( `* H5 f; m& ~+ U) e8 lStep 4 – Formulate Wash Buffers 0 n8 F+ a, F$ Z$ g, u  x
  Washing the Membrane  17 % K+ \% b" s. h) v
  Wash Buffers  17
3 r% X: [: g9 F  W5 V# NStep 5 – Detection Reagents ' T  ?; `- Z. S" F$ P  \' P' P
  Validated Primary Antibodies  18-19
& u. k+ F$ ~9 _+ |! X& j  Affnity-purifed Antibodies  20 & i* d' {- y' H; v! l
  Stabilized HRP Conjugates  21
) O0 E# _! Q$ t: ~! a  Thermo Scientifc DyLight   22
- ?( a" \6 \8 a% s. w   Fluor Conjugates   
0 v: N, E+ Y& @1 D  V, F  Conjugate Stabilizer Solutions  22 ! t! K% F& B2 v4 [
  Thermo Scientifc DyLight-labeled Highly  23 2 i1 y; o  g" Y# W  T$ G: c2 k, l6 S
   Cross-Adsorbed Secondary Antibodies
$ J; t2 T2 b- p  q; ~. X  Antibody Storage and Stabilizer Solutions  24
; ], q, Q+ ?2 [6 z! {+ D7 n  Secondary Antibody Ordering Table  25-26  
5 @6 y6 j; c% P+ r! @  Thermo Scientifc Clean-Blot IP  28-29 ( {$ |$ `1 g/ @. {9 i/ w3 T
   Detection Reagents   
) w  P  J9 |; ]  Protein A, G, A/G and L Conjugates  30
. i& ^2 a+ s$ [! o3 _( T" @  Thermo Scientifc NeutrAvidin, Streptavidin   31-33
. w1 s6 ~! K6 [- C/ r   and Avidin Conjugates% Q% a/ H5 h2 U8 K5 r6 C! B) ?
Step 6 – Enzyme Substrates
# P0 F5 Z5 n, x+ [. R5 ^Chromogenic Substrates  34-35
% |3 h; }# f# s2 tThermo Scientifc Chemiluminescent Substrates  36-43
+ X0 A2 I) ]5 R7 V5 Y. F8 v  Pierce ECL Substrate  37 " B# M) {  a; k4 x7 D+ j
  SuperSignal Chemiluminescent   38-42
" L8 |  h. D9 ~$ @   Substrates and Kits   
5 s4 n! A& y, X: P6 U0 z  Lumi-Phos™
5 Y3 @. \2 h' J* _* t5 X Chemiluminescent Substrate  43
5 k4 ~0 z% w% ], z$ I  Quick Reference Substrate Guide  43 : |- t: d5 l6 p; }; W( V
Fast Western Blotting Kit  44
, G/ I2 ~) A! Q3 |" }  a: L6 ZSpecialized Western Blotting Kits  45-48 ! N' Q3 e/ C, \2 K
  Thermo SuperSignal West Pico HisProbe Kit  45 3 c, R1 a/ `6 R$ c: k
  Pierce O-GLcNAc Western Blotting Detection Kit  46 0 R( f% _' c  X
  Thermo Scientifc DyLight 549/649  46-47  4 @. v% g$ n7 @$ `6 V5 ]  P. y
   Western Blotting Kit   
: b" R# u7 S, D$ H1 f  Thermo Scientifc DyLight 680/800 Near Infrared  47
( D& \  i7 j% M7 G   Western Blotting Kit   . L( ^% f: ]4 x$ L4 S# L
  Thermo Scientifc Active GTPase Pull-Down  48
) R# t1 d# D% w' Q( ?/ i4 S   and Detection Kits   
- C. Z. N) B% H  U( E  XFar-Western Blotting  49
  ]" K( R+ R+ U7 `In-Gel Western Detection  50-51
$ W1 ?4 ^) t2 X" c2 H  Thermo Scientifc Pierce In-Gel   51 : w. _7 X9 L: h( x) `
   Chemiluminescent Detection Kits6 o& D( X+ K1 ~1 a# s$ @4 I! E
Step 7 – Film
. ]& v! `+ j" a3 W2 }8 c+ P/ Z  Thermo Scientifc CL-XPosure Film  524 c" U3 E8 ]! V% z
Step 8 – Stripping Buffer 5 J. ?/ Q# ?* L- A/ ~/ Q
Optimizing the Signal-to-Noise Ratio  53-58 1 l, B- S0 t6 i
  Protocol for Stripping an Immunoblot  54 ! n" |) b' {; [. F
  Thermo Scientifc Restore  55 1 z3 f* x/ T" Y: y, D+ A
   Western Blot Stripping Buffers
( c+ T1 s0 y. Y% D  Thermo Scientifc Restore PLUS  56
8 C" A+ H3 u" v( |$ x4 ^   Western Blot Stripping Buffers
& p9 q0 |, t% Q; ^" D1 j  Thermo Scientifc Pierce   57-58   D+ E% }5 B9 U
   Background Eliminator
' x, i1 p% U! f" K) `% e* YTroubleshooting Guide  59-69
1 G+ i8 d# y1 J9 y2 z  Blotting with Chemiluminescence  59 8 n) x: R; g% N) ?* A" e: m; A
  Optimizing Antibody Concentration  60-63 * Y0 J( V) m$ [2 {* R! e
  Problem Guide  64-67 0 {1 Z( W' A1 K# k9 G" C# g7 w9 e
  Full-Length Western Blotting Protocol Using   68-69
  J8 H: q* j5 q. z   Chemiluminescent Substrates * S/ h- s* m" v& U
Recommended Reading  70
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4 z5 c2 @/ p7 J) J1 Y5 s( C[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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