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

dahui

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Introduction  1
6 R! W7 p. W5 X; \9 f# VWestern Blotting Overview  2-3
7 n7 _: v: x, u# ^" C: RStep 1 – SDS-PAGE
( m3 ?& u. v3 i' y  Thermo Scientifc Precise Protein Gels  4 ! N$ V1 ^. U0 ]& J1 m
  Molecular Weight Markers  5-6* R# D+ S6 f/ E8 G- P" A
Step 2 – Electro-Transfer ' k" @- m3 N! y9 }
  Thermo Scientifc Pierce Fast Transfer System  7
) ^: `/ |+ U0 {& e( s/ t  Transfer Buffers  8
' F! B9 F4 P" ?/ i& r5 W  Filter Paper for Blotting  8 ! P7 S  y8 _/ R, ~6 n4 r$ _% W
  PVDF and Nitrocellulose Membranes  9
6 N7 G5 b+ x2 i  z* j* x& `! u4 G6 Q  Thermo Scientifc Pierce Protein Stains   9  
: l( D3 a3 ^; H* W   for Membranes   
! D4 S' j" e0 }- i3 w9 D  Antibody Extender Solution NC  10 $ f" _( f1 m, q+ S/ E$ P/ j
  Western Blot Signal Enhancer  111 L" \' U1 o, x% C8 {0 Y1 d3 [
Step 3 – Blocking
  `7 b( ]3 g. E- u4 U+ L  Introduction  12
: U: L2 s! k3 t- |6 L& G+ ?) y  Blocking of Nonspecifc Binding Sites on  12 - ~( P9 S1 ^- _: s  A7 Q5 Y) {
   Transfer Membranes   . B. J, L3 {6 ]( o+ ~
  Blocking Buffer Optimization  12-13
1 J. p: P1 l! k+ k  Blocking Buffers  13-16
; N8 k, `4 g* D0 w) \, O- R& f: Y) s* _0 bStep 4 – Formulate Wash Buffers
7 k# f6 U" {- o' ]  Washing the Membrane  17 & T: A" }- J3 W# I& g7 `7 G
  Wash Buffers  17
+ H7 U7 a$ n% V  E' t# BStep 5 – Detection Reagents
) `; N$ {9 {. ?" N" V3 h  Validated Primary Antibodies  18-19
' l1 e1 Y; e0 z0 s! i1 `  Affnity-purifed Antibodies  20
% p8 M1 d) |0 K2 H  Stabilized HRP Conjugates  21 ; q# g: ~9 ^& u# B/ L8 P0 V
  Thermo Scientifc DyLight   22
, J1 l# t2 b( ~8 l9 s   Fluor Conjugates   
7 L" [7 n3 L& `. x  Conjugate Stabilizer Solutions  22
7 }% O" Z7 B) r  Thermo Scientifc DyLight-labeled Highly  23 ! y- ?3 r. }( v8 w
   Cross-Adsorbed Secondary Antibodies   u0 J, A  D: g
  Antibody Storage and Stabilizer Solutions  24 " C; _3 [5 H3 l
  Secondary Antibody Ordering Table  25-26  " P' }9 J* V: V0 J6 a
  Thermo Scientifc Clean-Blot IP  28-29 % B0 w7 Q) l* Q8 _8 V0 S
   Detection Reagents   
# g  ]* T4 e4 ]1 j0 O; ^  Protein A, G, A/G and L Conjugates  30 / C2 f: f$ a2 N5 B
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33
9 K- ?( K+ v: H2 K/ X; y8 [   and Avidin Conjugates
* g& k8 J7 Q7 P) WStep 6 – Enzyme Substrates ; }+ D) v" ]5 x* _0 M
Chromogenic Substrates  34-35 1 D, X7 L0 k, p: I
Thermo Scientifc Chemiluminescent Substrates  36-43 7 N- ]4 Y4 t1 Q! M+ t1 y' E% H. |9 y' n
  Pierce ECL Substrate  37
9 u- ?) V6 b, ]  SuperSignal Chemiluminescent   38-42 6 T% }: h/ Y4 K" F7 N
   Substrates and Kits   
! X; \  d: F9 s" N( Y: T  Lumi-Phos™
: C6 B0 U  x4 Q Chemiluminescent Substrate  43 & G" h" r8 j$ o; E0 M: l: j' ~7 P. x
  Quick Reference Substrate Guide  43 & J8 W; S; k2 [! R9 _5 _
Fast Western Blotting Kit  44 5 p) {8 B, u& M4 p! a7 N
Specialized Western Blotting Kits  45-48 ! q* Z% Q. c8 H4 @. S& t0 n$ b
  Thermo SuperSignal West Pico HisProbe Kit  45 1 J- P8 D% \0 d! [+ @0 b( X9 a
  Pierce O-GLcNAc Western Blotting Detection Kit  46
/ q$ V1 A2 q9 W( K  Thermo Scientifc DyLight 549/649  46-47  $ R( O3 K4 s# z6 q1 Y
   Western Blotting Kit   # w3 J2 _: T. w' Z+ W2 h
  Thermo Scientifc DyLight 680/800 Near Infrared  47 ! n: w3 |4 c& P7 J/ o  e& V3 q
   Western Blotting Kit   
- Z7 x5 \6 y* K# d7 |) Y  Thermo Scientifc Active GTPase Pull-Down  48
. E* g- N6 X  A& S9 ]& \) t) }6 j4 u   and Detection Kits   
" p( m9 x, }) j+ E; g3 uFar-Western Blotting  49 # [7 z* j/ \, w: ^8 K) k
In-Gel Western Detection  50-51 , b+ j/ u1 q# R6 V" U% t$ \) }+ U
  Thermo Scientifc Pierce In-Gel   51
/ j( F5 I& D5 L, Q6 `   Chemiluminescent Detection Kits
5 v2 q3 i5 n3 {+ H* X5 hStep 7 – Film 2 T( T; P8 g2 ]0 ]
  Thermo Scientifc CL-XPosure Film  52  v) k5 T% \' g" K5 I& a$ O
Step 8 – Stripping Buffer
6 Q3 d# s9 a$ q6 oOptimizing the Signal-to-Noise Ratio  53-58
9 w. E  g- i, B) U  Protocol for Stripping an Immunoblot  54 * ^8 P! x7 |& k2 ~+ P" g
  Thermo Scientifc Restore  55
% _: Y5 A8 }- {8 J# g4 L6 u   Western Blot Stripping Buffers   {; h- b' m0 O5 p
  Thermo Scientifc Restore PLUS  56 ; G6 X* p* `5 k4 t% v  |6 m& k: j
   Western Blot Stripping Buffers
2 \  c9 H6 y- h* B, P( Q6 l- y6 Z7 o% }  Thermo Scientifc Pierce   57-58 . ^7 D: m  L2 U* U1 W) a" q
   Background Eliminator% U& ~5 M! y6 [! R' Y( j
Troubleshooting Guide  59-69 ' e' M& b* W, p, O3 X+ f" D
  Blotting with Chemiluminescence  59 ; x9 b. Y8 x/ W7 r! J1 G6 s9 b
  Optimizing Antibody Concentration  60-63 * p! o6 L* D' Y7 A; f5 q3 A2 |
  Problem Guide  64-67
6 K. {6 N; F. A! g; Z. V  Full-Length Western Blotting Protocol Using   68-69 . w8 C+ h0 r  K
   Chemiluminescent Substrates 9 f* b% M! f; ]9 j. a4 ^
Recommended Reading  707 e) A+ G( [. W* A- N. [4 ~' o/ K5 P
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一枝独秀

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jiandong2001

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军医

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深海寂寞鱼

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sunyucheng

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daviddow

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mominglily

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