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

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6 f- O/ V( N( H" BIntroduction  13 g# v- z- M* r
Western Blotting Overview  2-3
: A/ t4 V* F& |; F: xStep 1 – SDS-PAGE
- M3 {& P7 r( V; ^% i. q+ I  Thermo Scientifc Precise Protein Gels  4 . B% C8 q7 z0 y) u4 C8 x
  Molecular Weight Markers  5-6
, [1 [" |1 R: ^  zStep 2 – Electro-Transfer
* Z: Y3 G7 w2 X4 \" D6 g  Thermo Scientifc Pierce Fast Transfer System  7
9 l  B4 {4 I' }' H/ x5 B. `5 b) D  Transfer Buffers  8
1 ~3 ~, R! y6 i3 E" S) o# J0 \  Filter Paper for Blotting  8
3 B# G: ^% z- W  PVDF and Nitrocellulose Membranes  9
, }' K! P4 t8 e6 }' B; r* Y) o  Thermo Scientifc Pierce Protein Stains   9  7 k4 ^9 G" Z% M/ J4 M. o( \
   for Membranes   ! _4 r6 U# d: q
  Antibody Extender Solution NC  10 1 V' c! m9 u/ ?9 S7 G
  Western Blot Signal Enhancer  116 V- ^2 H) [4 D" W  K
Step 3 – Blocking
/ F6 ~9 k8 Y+ R7 r  Introduction  12
" ?# U! O. E6 H3 {' G$ s  Blocking of Nonspecifc Binding Sites on  12 9 y% _, u; U3 M4 c
   Transfer Membranes   ' L$ [& Y* j5 U8 D$ V9 Y: y
  Blocking Buffer Optimization  12-13
7 R/ n; I1 d1 D  Blocking Buffers  13-16$ h. `6 X7 _, r
Step 4 – Formulate Wash Buffers . |+ U: u  U6 l- {, b- c
  Washing the Membrane  17
' e: E  ^! n& p8 L, W  Wash Buffers  172 m! P6 B9 \8 W" g
Step 5 – Detection Reagents
% z  b' }2 Y0 T1 x6 j* b+ H; B/ w  Validated Primary Antibodies  18-19
; a$ q/ C7 r# H2 ]" c) _4 l  Affnity-purifed Antibodies  20 9 y6 T$ R' H' j# _& q
  Stabilized HRP Conjugates  21 + H  V1 s7 C# {  b
  Thermo Scientifc DyLight   22 ( C* Y( c! ~! m' ?+ }& Q$ Y; r
   Fluor Conjugates   + B8 a! a5 [! Y* m
  Conjugate Stabilizer Solutions  22   \" @5 V1 d: B/ q
  Thermo Scientifc DyLight-labeled Highly  23
4 K- h, ^+ x. m, s- p   Cross-Adsorbed Secondary Antibodies " M7 N! I% r5 n) R* K* [/ D
  Antibody Storage and Stabilizer Solutions  24
& _, n+ H+ p4 |, L4 K; Y8 }  Secondary Antibody Ordering Table  25-26  
* ~' b# ]5 z9 }; E0 R/ J0 A6 X) I  Thermo Scientifc Clean-Blot IP  28-29 & g+ |/ [0 O8 w
   Detection Reagents   7 x- }( s- e0 B* v" N
  Protein A, G, A/G and L Conjugates  30
. k4 q! b4 |& r, x  Thermo Scientifc NeutrAvidin, Streptavidin   31-33
2 R1 ?1 l/ t: U1 Z   and Avidin Conjugates# G  Y. O- b9 O# f; ^
Step 6 – Enzyme Substrates
1 i3 g% D2 c/ I0 i3 A/ ^' NChromogenic Substrates  34-35 ; ]$ J, \7 {: o4 t% ], @* P- f; X
Thermo Scientifc Chemiluminescent Substrates  36-43 ! m+ ?; u8 h+ C' A2 O' y% w
  Pierce ECL Substrate  37
6 n& Q- w5 x; X6 \/ q  SuperSignal Chemiluminescent   38-42
! |# }( e4 v- H9 E' o1 H/ l1 \   Substrates and Kits   
  y+ A% b0 f5 Y" s- }3 f: @) r0 a* @  Lumi-Phos™* }9 E, G$ k7 N1 @) p  k
Chemiluminescent Substrate  43 - V6 m) T: `2 g
  Quick Reference Substrate Guide  43 9 h2 c/ y4 G8 C2 D( @
Fast Western Blotting Kit  44
' a; n) w3 ?4 ?  A6 g# e" PSpecialized Western Blotting Kits  45-48 , Z. o/ w% M0 w- E. {; X- u
  Thermo SuperSignal West Pico HisProbe Kit  45 2 T( T. {. h+ l1 n: f5 ?
  Pierce O-GLcNAc Western Blotting Detection Kit  46
. [- p& F) F5 k  Thermo Scientifc DyLight 549/649  46-47  ) s% U( Q* n' X; T9 H* C
   Western Blotting Kit   
/ x+ y' J3 p% n  Thermo Scientifc DyLight 680/800 Near Infrared  47
( \/ b) x# e$ T+ Y1 Y/ R   Western Blotting Kit   4 o7 b/ L* C9 U  W- c
  Thermo Scientifc Active GTPase Pull-Down  48 * h( O7 e" R* E2 R; R! D1 x4 Q
   and Detection Kits   
2 M0 i# d2 s3 c6 H- C5 pFar-Western Blotting  49
* O$ m* m$ d9 {/ {. A% mIn-Gel Western Detection  50-51 9 @+ a' ]7 K/ d+ X
  Thermo Scientifc Pierce In-Gel   51 $ @- f1 D- M  n/ M9 c9 y& Z
   Chemiluminescent Detection Kits+ c5 F; ^! i( m
Step 7 – Film
# J, ?: v, p7 z9 @" h1 c  Thermo Scientifc CL-XPosure Film  52
, p9 i. B; ~0 u# d8 N/ OStep 8 – Stripping Buffer
1 |; x; q& Q3 E" AOptimizing the Signal-to-Noise Ratio  53-58 # \9 r( o# s* o" w- z) X7 C: G
  Protocol for Stripping an Immunoblot  54
$ C6 u% n) L) t9 b8 e  Thermo Scientifc Restore  55
4 {% {8 n/ V6 ]4 ?3 R! G0 I3 P   Western Blot Stripping Buffers
7 L( E% M4 p8 Z2 @% H! m  Thermo Scientifc Restore PLUS  56
* ?8 O9 n" @! X, m1 ?   Western Blot Stripping Buffers
* [7 X/ t$ r: F6 P  Thermo Scientifc Pierce   57-58   I* A" F  g; ~" I& f0 ^
   Background Eliminator" h  e) U8 s7 N
Troubleshooting Guide  59-69   r8 s! e$ A) \) Z9 u$ \7 I
  Blotting with Chemiluminescence  59 ; z/ Z& X7 d+ y0 b1 o
  Optimizing Antibody Concentration  60-63   w1 P# {- q" M" x* T
  Problem Guide  64-67 1 p. v( Y/ M3 X7 k
  Full-Length Western Blotting Protocol Using   68-69
. C1 U" n5 a. P8 ]   Chemiluminescent Substrates + K, _. D' l- R, h% h
Recommended Reading  70
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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

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daviddow

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mominglily

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