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

dahui

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  ~4 n, l8 ?2 b3 V; _2 h; _+ z& r/ AIntroduction  1
9 C$ C6 g$ r3 }. l  f4 ~0 [Western Blotting Overview  2-3- H! j( L, n7 Z: P. \6 J. c
Step 1 – SDS-PAGE 5 Q' R% `" t7 l9 [$ c3 x) s, E$ \) b
  Thermo Scientifc Precise Protein Gels  4 6 I- a; M6 Z! D1 O
  Molecular Weight Markers  5-6
3 ]7 R" }- g) s5 j8 @Step 2 – Electro-Transfer ( J7 F0 G/ f6 E) Y. U
  Thermo Scientifc Pierce Fast Transfer System  7
( e1 r" Y6 c; f) l: X# A  Transfer Buffers  8
" o4 r0 Q6 v9 H( t% d) J' P  Filter Paper for Blotting  8
. _) n8 O5 {7 k/ z6 D# s4 g" |) e+ a  PVDF and Nitrocellulose Membranes  9 ! K1 e3 E7 g/ ^& O( s
  Thermo Scientifc Pierce Protein Stains   9  
  e1 ^  m5 h, {3 S* v- k! d) y2 p   for Membranes   
" p+ A. c3 A% R7 H  Antibody Extender Solution NC  10 6 U4 e2 T* _  z
  Western Blot Signal Enhancer  11$ c+ p* _! k1 y2 q; x3 Z) H' b
Step 3 – Blocking
% X+ `1 x5 A0 u  p- a" |& b2 w  Introduction  12 ( u/ Q, Y+ X( u# P  |
  Blocking of Nonspecifc Binding Sites on  12
+ M# _! t; e. k, M/ h  i) ]   Transfer Membranes   
4 M* j0 h, w8 Y3 g. I  Blocking Buffer Optimization  12-13
# _& \9 t% F) o2 t  Blocking Buffers  13-16& T. \6 |0 s0 @$ H
Step 4 – Formulate Wash Buffers
* C! [9 g( n  Q  Washing the Membrane  17 4 G3 S' o, O( X4 |9 n
  Wash Buffers  17
+ z* u% c" H: \% r$ {Step 5 – Detection Reagents
: V0 v" j1 j. x7 L. }  Validated Primary Antibodies  18-19
7 x, d5 _$ V; {  Affnity-purifed Antibodies  20
- M! _! l, ]: ^- U: z5 ]# t  Stabilized HRP Conjugates  21 5 X  B6 v8 T2 H
  Thermo Scientifc DyLight   22 ( D/ \+ r. l6 X& L- e! ~: o
   Fluor Conjugates   ( j$ m/ H7 ?1 e9 F
  Conjugate Stabilizer Solutions  22 9 V& m9 i$ N4 f4 H) T8 d$ K+ V
  Thermo Scientifc DyLight-labeled Highly  23 1 z. Z4 T" `: d0 p3 ?" T- K
   Cross-Adsorbed Secondary Antibodies
% N# F" ]- t: I9 A: ^  Antibody Storage and Stabilizer Solutions  24 $ J- z9 Z; h& C8 X1 T
  Secondary Antibody Ordering Table  25-26  
$ q( o, p6 I1 }4 K  Thermo Scientifc Clean-Blot IP  28-29 / H0 o, i: u* z" @! X+ c& W( M  N( @+ S
   Detection Reagents     X9 i5 ~0 r' L( }5 G2 _4 e
  Protein A, G, A/G and L Conjugates  30
1 t# g& [1 s  K1 V4 f5 T" m  Thermo Scientifc NeutrAvidin, Streptavidin   31-33 9 s4 C: ?5 A  |5 w* ]4 i
   and Avidin Conjugates
1 h- ]( m! m* ]Step 6 – Enzyme Substrates
0 Y. y% v2 L: j* b0 F( m: x) o* FChromogenic Substrates  34-35 4 z& s0 Y! L! I) u; b
Thermo Scientifc Chemiluminescent Substrates  36-43
; m2 U9 Z$ i6 l0 y; v0 O. P  Pierce ECL Substrate  37
, V- t1 ?  A$ A3 g  j0 v7 N  SuperSignal Chemiluminescent   38-42 6 Q' a4 n3 v' {0 w" t* R
   Substrates and Kits   4 Z& T, I6 z: V( [# X& V; ?: D
  Lumi-Phos™% m. i( T. E, c: d% b
Chemiluminescent Substrate  43
- s+ |3 o  y  P- l0 h/ O  Quick Reference Substrate Guide  43 5 Q4 }0 j, i( [1 {/ M
Fast Western Blotting Kit  44 / o$ u9 s) m1 J7 A. F' R7 w0 X1 o
Specialized Western Blotting Kits  45-48 - n; @! g4 l: C- @
  Thermo SuperSignal West Pico HisProbe Kit  45
$ T" k- @# J6 d  Pierce O-GLcNAc Western Blotting Detection Kit  46 7 ]: M  q2 U5 r1 Q7 e
  Thermo Scientifc DyLight 549/649  46-47  
/ v- z  w5 G9 \6 y7 K   Western Blotting Kit   
, i7 @& ^3 R" T, _0 ^) d  Thermo Scientifc DyLight 680/800 Near Infrared  47
: K6 I! ^& Q" a( _* E8 _  P2 S$ z5 S   Western Blotting Kit   % s5 l. b6 u4 P6 O7 X& }/ [* e
  Thermo Scientifc Active GTPase Pull-Down  48
8 d/ D% W6 r- ~; y. m   and Detection Kits   
8 G+ ^2 a- k3 y& tFar-Western Blotting  49 8 q6 v' G5 C7 Z6 C: w/ c
In-Gel Western Detection  50-51 6 g- @" _. z& u4 y/ x& z7 Q
  Thermo Scientifc Pierce In-Gel   51 2 L9 ~  i: l/ E6 u' u
   Chemiluminescent Detection Kits
9 X& S( u. N% k1 x7 FStep 7 – Film
- D( D! |6 \/ `9 b. A/ R9 M  Thermo Scientifc CL-XPosure Film  52
  y- U; I; f! @2 e3 SStep 8 – Stripping Buffer
( ?1 E/ }" g) F) ]Optimizing the Signal-to-Noise Ratio  53-58
3 e! V. k6 p( F5 g0 M/ r  Protocol for Stripping an Immunoblot  54 / A% z, n6 n# a+ `/ h) Y$ n% @
  Thermo Scientifc Restore  55 " [5 P# E! L* v3 M( g4 c
   Western Blot Stripping Buffers 7 N7 C! h- v8 Z- s
  Thermo Scientifc Restore PLUS  56
5 Z4 R  [6 H- F4 ~4 ~0 X9 e   Western Blot Stripping Buffers : M; m* g/ C, q! K# V/ r
  Thermo Scientifc Pierce   57-58
# `' l% N# `7 h$ s7 W   Background Eliminator
4 o( U3 r6 g; ~Troubleshooting Guide  59-69 . e0 d1 ]* }1 g# ^( n% m! p
  Blotting with Chemiluminescence  59
! o+ w6 \1 g9 K  G+ g7 S! t  Optimizing Antibody Concentration  60-63
' a4 W9 b  b' v8 c4 h  Problem Guide  64-67
! C* h7 r5 m& x# d4 r  Full-Length Western Blotting Protocol Using   68-69
# {* M# t* C8 @0 G% W$ L, g/ l   Chemiluminescent Substrates & ~3 q! i1 [( f) m3 E4 j1 k
Recommended Reading  70
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' ?$ ^3 u' @" }5 B4 L[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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