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

dahui

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Introduction  12 `  z$ d( i" K
Western Blotting Overview  2-3
" |. J  P# m7 LStep 1 – SDS-PAGE
! f* ]6 r+ T9 H3 T2 \2 C  Thermo Scientifc Precise Protein Gels  4
0 t  _) T' V2 [% D, L. q  Molecular Weight Markers  5-6
; n( @* q$ P# T+ n2 d# sStep 2 – Electro-Transfer
4 ^9 r4 u, g: w2 L  ^- K1 X6 ~! J2 f  Thermo Scientifc Pierce Fast Transfer System  7
0 @6 A; |9 U  H. F& }. E+ u* f- l  Transfer Buffers  8 " H3 S" `5 H- W3 o4 `  s
  Filter Paper for Blotting  8
: W& e/ [1 u1 t  PVDF and Nitrocellulose Membranes  9
2 j- q9 q7 L5 r7 u  Thermo Scientifc Pierce Protein Stains   9  ( Q$ L# d) \) L8 y: ^
   for Membranes   ; o  T: a" U7 P
  Antibody Extender Solution NC  10 0 o) U; g, l- C7 j! E  L
  Western Blot Signal Enhancer  11
1 s6 z, t$ R3 Q2 BStep 3 – Blocking + C; i& m5 K) x. U! D
  Introduction  12
% b1 c4 L5 V3 U. a# w' m  Blocking of Nonspecifc Binding Sites on  12 . a7 a  |7 D  _: w; v  u* T- S
   Transfer Membranes   , m5 C/ ]  V' E$ C# t
  Blocking Buffer Optimization  12-13
5 a; Q! Q' E7 f" |5 r; Z  Blocking Buffers  13-167 L8 K5 m; d1 g. ]" R. @: Z; T
Step 4 – Formulate Wash Buffers & \* \1 A1 K& e6 |) ?' C. M; D
  Washing the Membrane  17 : c' Q2 J9 t# b# Q9 G0 E
  Wash Buffers  17
; t% e3 _' @. D. h) h; aStep 5 – Detection Reagents
- x2 ^9 ]- p) l8 {, |8 X9 \- X: D) i  Validated Primary Antibodies  18-19 4 _6 V: C8 Y/ H; x7 a5 i
  Affnity-purifed Antibodies  20
6 j. a( S6 B7 e- T4 F. F0 e  Stabilized HRP Conjugates  21 - u2 i9 D% z* z( a, P- u7 k1 |
  Thermo Scientifc DyLight   22 ! p, z5 {0 W. O+ a' a3 T
   Fluor Conjugates   , j4 A5 `  i9 ?( a- p
  Conjugate Stabilizer Solutions  22
/ [: o1 ^" k2 M7 b" X! }  Thermo Scientifc DyLight-labeled Highly  23 5 i! e2 q# o: L6 R+ T: L& p
   Cross-Adsorbed Secondary Antibodies + C( r( V$ r' Z4 P# j
  Antibody Storage and Stabilizer Solutions  24
4 t7 D6 ]& D/ n3 l  Secondary Antibody Ordering Table  25-26  % [% ?8 T! D0 C- M7 H; f
  Thermo Scientifc Clean-Blot IP  28-29
& }/ ?5 D' \5 ?$ F   Detection Reagents   + l& p7 c: z3 G& j/ a1 ^, u
  Protein A, G, A/G and L Conjugates  30 & U3 X% J. F. t# }$ y
  Thermo Scientifc NeutrAvidin, Streptavidin   31-33
: H+ K3 g6 R6 v' w. S4 ^   and Avidin Conjugates! i8 q/ a( \0 h% a
Step 6 – Enzyme Substrates 0 m2 H6 j/ A/ i9 q7 [: f) |+ r! X
Chromogenic Substrates  34-35 * m! }7 h" r9 @* `7 B. p  o+ {  k2 {
Thermo Scientifc Chemiluminescent Substrates  36-43 ! S: P; r) y2 e; \* B, V
  Pierce ECL Substrate  37 * M* `' {3 p9 n% b
  SuperSignal Chemiluminescent   38-42 3 z7 k  r! t4 l( \- g: z
   Substrates and Kits   
) d3 ]$ I$ e& h/ f7 X" c! c  Lumi-Phos™
0 u, U; z/ j4 j) s8 t Chemiluminescent Substrate  43 - H* C3 ]$ D8 j$ w9 L: E+ P
  Quick Reference Substrate Guide  43 ; ]2 {7 ~% Y3 x7 H/ c, V. g
Fast Western Blotting Kit  44
  C$ b: C5 u4 k5 o; B0 Y+ ?Specialized Western Blotting Kits  45-48
4 }+ v! |5 Y' j: N8 _! S  Thermo SuperSignal West Pico HisProbe Kit  45
$ \7 r0 S9 Q$ j9 A  Pierce O-GLcNAc Western Blotting Detection Kit  46 + ?; u/ v* {  p0 k. J8 {  p
  Thermo Scientifc DyLight 549/649  46-47  ; r8 g5 l# \( G7 [
   Western Blotting Kit   ! }$ }' d# O# P3 D
  Thermo Scientifc DyLight 680/800 Near Infrared  47 - k' A/ g7 D( {* F( Y
   Western Blotting Kit   
7 H! O; k+ z8 s  Thermo Scientifc Active GTPase Pull-Down  48 9 k  D0 ^/ d2 y6 T! e
   and Detection Kits   / W  ]2 B3 n: m- j7 t
Far-Western Blotting  49
. B' d" I7 |$ V, hIn-Gel Western Detection  50-51
9 h. _! L8 i. W" Q  Thermo Scientifc Pierce In-Gel   51
( q% u( p( y5 h% j) M% Y1 I   Chemiluminescent Detection Kits$ J9 y0 i: r7 g8 u# P
Step 7 – Film 2 R" r3 j$ i+ G( w* \
  Thermo Scientifc CL-XPosure Film  52
! ?5 N7 s) w( ?* j: DStep 8 – Stripping Buffer
) b5 p4 ^' m3 oOptimizing the Signal-to-Noise Ratio  53-58 : t+ E5 T7 f9 z+ H9 j
  Protocol for Stripping an Immunoblot  54
2 `- W* [- G$ i; @& c0 J  Thermo Scientifc Restore  55 8 p* b5 d  u6 w9 C, w% o2 C
   Western Blot Stripping Buffers # c7 s0 j) Z' O* y
  Thermo Scientifc Restore PLUS  56 ) ^1 b" x9 ~; M' [7 e$ p
   Western Blot Stripping Buffers
! X- g7 U. C9 G  K1 f- F" P  Thermo Scientifc Pierce   57-58
- `/ c4 ?8 c" i) |8 `# O   Background Eliminator' D5 X! Y- i4 K# R9 i  F
Troubleshooting Guide  59-69
7 Q! H. t2 d! ~7 f) P0 O0 |  Blotting with Chemiluminescence  59
  U' x* `: |5 ]  Optimizing Antibody Concentration  60-63
0 c) p, g3 _5 d. ]' P7 Y, @" C4 C  Problem Guide  64-67
9 U. {' I4 @) D: ?  Full-Length Western Blotting Protocol Using   68-69
" [2 c$ j5 N# k& @9 Y   Chemiluminescent Substrates 9 |* f! Q( s( T3 o; \9 z3 k
Recommended Reading  709 E2 r5 W  x1 x4 q

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一枝独秀

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jiandong2001

好久没有来这里了！谢谢分享

军医

谢谢分享！

深海寂寞鱼

好资料 学习ing

白衣小彩

学习学习

sunyucheng

看看

daviddow

thanks for sharing it

mominglily

谢谢

qlwj

新手上路，能发到我的邮箱吗？我看不到内容ql_tjgk@hotmail.com