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

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Introduction  1' x/ G0 D! }" {
Western Blotting Overview  2-3
: ?% n4 ~- G  ^6 \- m' \; [+ w# A- a6 IStep 1 – SDS-PAGE ( a5 y, ~/ B1 ?$ M6 {
  Thermo Scientifc Precise Protein Gels  4
4 w3 n* G; V$ e- f; ~  Molecular Weight Markers  5-66 d9 S/ H" B( v4 v5 S0 R6 S
Step 2 – Electro-Transfer 3 i/ \( {2 M; e
  Thermo Scientifc Pierce Fast Transfer System  7
7 E6 S3 e% ^0 ~2 o% k  Transfer Buffers  8 ; h7 F+ w  p/ g8 W# M( P# J" ?
  Filter Paper for Blotting  8
- T) I1 H  N1 m5 @4 k' w& J. J  PVDF and Nitrocellulose Membranes  9
6 n/ d) k! [8 }5 n/ I% M) b/ I  Thermo Scientifc Pierce Protein Stains   9  
6 \# ~4 S$ Q1 N: S$ |/ F0 z   for Membranes   ! ~) G& S' W, J- X! d
  Antibody Extender Solution NC  10 , j5 R# |+ N* j9 [
  Western Blot Signal Enhancer  11
+ w0 R3 |$ x6 aStep 3 – Blocking 8 t5 j* j5 V& @  |: e% S7 E  q$ T
  Introduction  12 . \( C5 z  g/ K
  Blocking of Nonspecifc Binding Sites on  12 # o) w0 H, W: g
   Transfer Membranes   
& w7 R/ U+ h. e% s2 g1 n  Blocking Buffer Optimization  12-13 5 c7 u& H+ x0 R" u2 R
  Blocking Buffers  13-16  s6 A5 L" s9 Y9 F7 O
Step 4 – Formulate Wash Buffers % c1 k3 e4 u- P' C* E- a
  Washing the Membrane  17 - Q* y$ t" @3 P# f: j
  Wash Buffers  17! O( a% Y! X9 W; D8 c9 ?
Step 5 – Detection Reagents
* ^, @2 A2 C9 v: g' \  Validated Primary Antibodies  18-19 ' N3 [7 w. O# T: a: S& G) t+ z- e$ [
  Affnity-purifed Antibodies  20 ; N  L" ]+ y$ K
  Stabilized HRP Conjugates  21 % B" l$ @$ e7 s! Y7 x" P) W
  Thermo Scientifc DyLight   22 2 x4 K$ q+ }+ m8 }; z' I
   Fluor Conjugates   
% u& M4 e" h; P8 c! J# G  Conjugate Stabilizer Solutions  22
. q9 z1 ~4 Y; \+ |8 o  Thermo Scientifc DyLight-labeled Highly  23 , A! N- c* o* Z' |" A
   Cross-Adsorbed Secondary Antibodies
5 S$ I8 N" w: D( m$ O0 ~2 V  Antibody Storage and Stabilizer Solutions  24
( E# G; ~% N8 p) x: e  Secondary Antibody Ordering Table  25-26  
" G* v: U! t- X7 S3 f' {. Z  Thermo Scientifc Clean-Blot IP  28-29 * v' L3 P7 A/ O
   Detection Reagents   0 \9 a9 q0 |( h; b
  Protein A, G, A/G and L Conjugates  30 / K1 {) @" L0 _6 q3 y9 q
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 . I% u" u! P' q" F
   and Avidin Conjugates$ z( V7 a' k% O2 o; r2 P- M
Step 6 – Enzyme Substrates / [) Z! k8 M: U) w: i, J$ w
Chromogenic Substrates  34-35 ) N! `+ q/ |* H$ u; x, E5 _3 M  I6 Z
Thermo Scientifc Chemiluminescent Substrates  36-43
( C2 P" k2 L& S4 B0 A. k9 Z  Pierce ECL Substrate  37 ' {3 X) a3 y3 r! o+ c" Z. H  W/ L! n
  SuperSignal Chemiluminescent   38-42 7 J3 W, W9 B+ V# X
   Substrates and Kits   
. Z# @, V& h4 G  Lumi-Phos™
/ N! V3 w/ I3 s7 T  e/ k Chemiluminescent Substrate  43 * e3 Z, @: W/ l  {# U) p
  Quick Reference Substrate Guide  43
) r* a* z) Q6 ?3 |0 _) C: X* O, o1 U' aFast Western Blotting Kit  44
, U9 \* Y, j! b# K& a/ {! {' iSpecialized Western Blotting Kits  45-48
7 X# z0 O! R, h  E. K( s1 S  Thermo SuperSignal West Pico HisProbe Kit  45
( j4 E5 J3 J( k  Pierce O-GLcNAc Western Blotting Detection Kit  46
+ v& V2 R- O' q  Thermo Scientifc DyLight 549/649  46-47  
/ i2 V. c/ ^3 `   Western Blotting Kit   
. Z9 [) y* m8 E3 [4 h0 z5 Q  Thermo Scientifc DyLight 680/800 Near Infrared  47
4 |% u$ j) ^: C9 P  G   Western Blotting Kit   ( C5 t! D- Y4 z8 m( h5 r
  Thermo Scientifc Active GTPase Pull-Down  48 1 y( C% O. V2 t, _) T5 ?, M" ~) j
   and Detection Kits   
) z+ u) K) p2 [% `4 b# aFar-Western Blotting  49
" W/ ?* `$ o* u* ^7 g1 L  A: ~In-Gel Western Detection  50-51
1 b4 m! S# ?" P$ s7 r: N; j# b  A  Thermo Scientifc Pierce In-Gel   51
, `9 }5 a/ U! L+ |. j( v0 m  Y   Chemiluminescent Detection Kits
/ [' q/ r* `+ ZStep 7 – Film
; h1 _1 v+ G6 q$ E5 C/ `3 o  Thermo Scientifc CL-XPosure Film  52
- a, ~6 G9 ]  [Step 8 – Stripping Buffer
- Y# x7 }1 C! uOptimizing the Signal-to-Noise Ratio  53-58 % L8 `# @7 ?0 j& o% O; O& C/ D6 ?
  Protocol for Stripping an Immunoblot  54
# V/ l& z/ R1 r1 e  Thermo Scientifc Restore  55 - n9 n& K0 S$ i) u- n, K. U3 v
   Western Blot Stripping Buffers 2 W  W4 \, K( w8 Z, b' @
  Thermo Scientifc Restore PLUS  56 . \( S2 l: F! S8 h' f
   Western Blot Stripping Buffers 2 U$ G, ]; i8 B5 {3 q: Q. q3 K# z6 m
  Thermo Scientifc Pierce   57-58
7 g- G! X: o; C; K/ m: q- s, Z   Background Eliminator7 z2 U5 \; {6 J" ^; L
Troubleshooting Guide  59-69
  B; h; q/ i3 Q, O: o6 s% C( ]  Blotting with Chemiluminescence  59
3 u( ?9 z% ~8 x5 P+ F  Optimizing Antibody Concentration  60-63
# R- K4 H! N1 x) k: i, k  N  q  Problem Guide  64-67
: X2 m/ {0 n1 ]3 l  Full-Length Western Blotting Protocol Using   68-69
1 O4 q5 v3 }: W. e! D5 r   Chemiluminescent Substrates
% F; p! G+ D+ VRecommended Reading  70, ^) `% f8 }: ?; p$ V2 w

6 l  {; M  `! p+ Q[hide][/hide]

碧湖冷月

原来是这样  

小小C

原来这样也可以  

leeking

支持一下吧  

甘泉

内皮祖细胞

pcr

想都不想，就支持一下  

舒思

呵呵 大家好奇嘛 来观看下~~~~  

海小鱼

写得好啊  

lalala

呵呵，找个机会...  

feixue66

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