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

dahui

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0 F8 [: ~0 n( N) y% lIntroduction  1
) |( E6 d( a1 O" XWestern Blotting Overview  2-33 l$ s2 f. T0 z8 I0 T/ \
Step 1 – SDS-PAGE
  _$ z1 ~5 q$ `6 U  Thermo Scientifc Precise Protein Gels  4
, \7 C% Q$ U) t' r5 {+ `6 ^1 Z  Molecular Weight Markers  5-6
1 g$ ^" ]6 u; ]$ L7 j$ q& |" nStep 2 – Electro-Transfer
) R& v. v; x! r& p: [8 o2 J7 T  Thermo Scientifc Pierce Fast Transfer System  7 2 X% [* m; n# T- r7 O1 g1 [8 }
  Transfer Buffers  8
* t8 R$ t: z' F$ h9 u1 c  Filter Paper for Blotting  8
& D& X4 t* A$ Z6 {. ?8 n  PVDF and Nitrocellulose Membranes  9 8 J" ~: Z' C, H2 @
  Thermo Scientifc Pierce Protein Stains   9  
  m6 |% E" H1 z# @+ f   for Membranes   5 B7 W+ E3 Z; Q0 e. N
  Antibody Extender Solution NC  10
6 P5 v: k" \6 G: i) V  Western Blot Signal Enhancer  118 P+ D3 s/ I6 m  k: K' k. U6 o; s
Step 3 – Blocking & K$ W. B- G  r/ E' T1 o, P
  Introduction  12 3 X; U7 n% W, P
  Blocking of Nonspecifc Binding Sites on  12 & r: P# i! b$ t/ q: K% K/ O
   Transfer Membranes   
" n0 K7 R# u& ~: C- Q* w5 P9 Q) |  Blocking Buffer Optimization  12-13 % O6 r/ F) U: {+ i% U  B; [4 @% T
  Blocking Buffers  13-166 b3 A) J6 H5 k1 {
Step 4 – Formulate Wash Buffers
! D3 t; m2 u6 r. a$ E. J" m  Washing the Membrane  17 ' l5 E  r" T  r% h6 Q; b
  Wash Buffers  17: \+ E5 H* @$ ]8 y* u, g
Step 5 – Detection Reagents 8 ]7 F% f6 @, i) O, T" G
  Validated Primary Antibodies  18-19
8 Y/ p0 k2 \2 a& i! \* Z  Affnity-purifed Antibodies  20 8 I* I/ t+ K' S$ t# y
  Stabilized HRP Conjugates  21
1 w2 Y/ J' E; ~4 Y# f4 g  Thermo Scientifc DyLight   22
- J6 I' d3 L3 \. D5 k   Fluor Conjugates   
& q  p: z- M4 H0 r. S  Conjugate Stabilizer Solutions  22 5 p, u& y2 O  |9 p) m: W
  Thermo Scientifc DyLight-labeled Highly  23
/ ]* F1 N* A( s) B9 E4 e. F   Cross-Adsorbed Secondary Antibodies : l% d! g! I; g4 T: I
  Antibody Storage and Stabilizer Solutions  24 ; Q$ f# M$ k0 D$ t0 a4 u4 a
  Secondary Antibody Ordering Table  25-26  
# F2 S' C9 v& j9 e% F& Z  Thermo Scientifc Clean-Blot IP  28-29 ) J8 u! L, \3 x4 X. x+ j, y  h- E
   Detection Reagents   
/ `0 M( r+ D! U; i4 A% b  Protein A, G, A/G and L Conjugates  30 : i0 S- q% i) ]5 m
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 2 [- P. b# K: m! i0 A: j3 W' Q
   and Avidin Conjugates
/ G+ O- p7 H+ N0 f1 o, E! g9 T! E. ~' jStep 6 – Enzyme Substrates $ k% O' F# L: e/ n, o. A
Chromogenic Substrates  34-35
1 B; V# n0 ]* c# V+ }/ uThermo Scientifc Chemiluminescent Substrates  36-43
8 I. t/ p5 l' Q, K) F& B' c7 N  Pierce ECL Substrate  37 ' h0 b* G! N3 T) J$ B
  SuperSignal Chemiluminescent   38-42
3 j9 E' O# e1 W& b  \   Substrates and Kits   
2 p6 D- w' V0 Q) w  c  N+ h# _7 |5 q  Lumi-Phos™
! e, K. [. \) X* D4 A Chemiluminescent Substrate  43
9 o1 ?1 l1 k. T" `  Quick Reference Substrate Guide  43 $ H) |1 o% N# H, u7 t, w
Fast Western Blotting Kit  44 9 c$ X8 R' _) {, ?1 L$ |- z
Specialized Western Blotting Kits  45-48 ) Z  L7 m0 l0 j" F+ G! t& k
  Thermo SuperSignal West Pico HisProbe Kit  45 ! q( K6 Y: c: h& z# \1 c: f2 [$ o
  Pierce O-GLcNAc Western Blotting Detection Kit  46
; j) ]5 M% R3 g3 \1 ?# V- C  Thermo Scientifc DyLight 549/649  46-47  - V4 d. j7 {" ]
   Western Blotting Kit   
% |( U3 ?5 r3 E' {! j$ z5 k  Thermo Scientifc DyLight 680/800 Near Infrared  47
) a: N6 b, p; ?& P& N   Western Blotting Kit   ; q7 ^4 y' e  K, I; S/ o" N; T
  Thermo Scientifc Active GTPase Pull-Down  48
6 G" L$ _5 a3 h( ^5 f5 z( J   and Detection Kits     T4 E# }1 O/ Z8 Y9 g. f
Far-Western Blotting  49
3 \/ p/ `: O: D- G0 E- H, M8 F" PIn-Gel Western Detection  50-51
# s7 E" g5 d( P9 D0 ]3 B, i  Thermo Scientifc Pierce In-Gel   51 & s1 ?1 ?7 r6 X
   Chemiluminescent Detection Kits
+ c, j; n1 v# M: w$ MStep 7 – Film
6 i, {. n8 h: f  Thermo Scientifc CL-XPosure Film  520 E. `- O# J3 ]
Step 8 – Stripping Buffer
$ n' w9 u" }( {# G# QOptimizing the Signal-to-Noise Ratio  53-58
& Q/ Q" H# f3 y2 j  Protocol for Stripping an Immunoblot  54 0 w3 o3 \7 o. A8 E3 a
  Thermo Scientifc Restore  55 / f4 f% v- Q  ?- ?
   Western Blot Stripping Buffers
& X! Y0 r6 I5 E& e* J0 q  Thermo Scientifc Restore PLUS  56 7 q& Z, x3 a; ^) p0 H: N8 P
   Western Blot Stripping Buffers
" W  z' Y0 Z8 E0 w1 a, X  Thermo Scientifc Pierce   57-58
/ V# k* n5 K; g& H) L" J   Background Eliminator
5 \& D$ |0 f- W* S9 ZTroubleshooting Guide  59-69
0 ?% V9 u) ]6 y* z1 W  Blotting with Chemiluminescence  59 - b( O* J( m- K: U, D0 U1 q
  Optimizing Antibody Concentration  60-63 8 Y' P/ e- _4 U' ]6 S
  Problem Guide  64-67 . s8 w. w8 `! e/ P1 ]3 o, z" R
  Full-Length Western Blotting Protocol Using   68-69 & v' [1 H& D5 J! x
   Chemiluminescent Substrates & S% m, I2 u) C. u9 f0 I
Recommended Reading  70* b5 z  `  `) @  h4 ?
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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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白衣小彩

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sunyucheng

看看

daviddow

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mominglily

谢谢

qlwj

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