试剂配方

WB实验相关试剂配方

RIPA lysis buffer For 1000 ml
50 mM TrisHCl, pH 7.4 (1 M stock) 50 ml
150 mM NaCl 8.76 g
1% Triton X-100 10 ml
0.5% Sodium Deoxylcholate 5 g
0.1 % SDS 1 g
10 mM NaF 0.41 g
1 mM EDTA (0.5 M stock) 2 ml
Add ddH₂O to 1000 ml  
Adjust to pH 7.4  
Add PMSF to 1 mM and other protease inhibitors immediately prior to use. 
4X SDS sample buffer  
150 mM Tris•HCl (pH 7.0) (1M stock) 15 ml 
25% Glycerol 25 ml
12% SDS 12 g
0.05% Bromophenol Blue 0.05g
6%  β-mercaptoethanol
 6 ml
Add ddH₂O to 100ml, aliquot and store at -20°C  
1X TBST  
20 mM Tris-base  2.42 g
150 mM NaCl  8.76 g
50 mM KCl  3.73 g
0.2% Tween-20  2 ml
Adjust pH to 7.6  
Add ddH₂O to 1000ml
Wet transfer buffer  
25 mM Tris-base 3.03 g
192 mM Glycine 14.4 g
20% Methanol 200 ml
Add ddH₂O to 1000ml  
Semi-dry transfer buffer  
48 mM Tris-base 5.81 g
39 mM Glycine 2.93 g
0.0375% SDS 0.375 g
20% Methanol 200 ml
Add ddH₂O to 1000ml
Separating gel (ml, total 10 ml)        
MW of target  protein (kDa)
 80-200 35-100 25-60 20-40
Gel percentage 8% 10% 12% 15%
ddH₂O  2.1  1.5  0.8  0
30%  Acrylamide
  2.7  3.3  4  5
2x Separating  buffer
  5.0  5.0  5.0  5.0
10% APS  0.1  0.1  0.1 0.1 
TEMED  0.01  0.01  0.01  0.01
Stacking gel (ml) 4 ml 6ml 8ml
MW of target  protein (kDa)
 - - -
Gel percentage 4% 4% 4%
ddH₂O 1.4 2.1 2.7
30% Acrylamide 0.5 0.8 1.1
2x Stacking buffer 2.0 3.0 4.0
10% APS 0.04 0.06 0.08
TEMED 0.004 0.006 0.008
2x Separating Buffer Recipe (makes 1000ml)  
Tris HCl (pH 8.8) 90.8 g
SDS 2.0 g
Dissolve compounds thoroughly. Adjust pH slowly to pH 8.8 with concentrated HCl, then add ddH2O to 1000ml.
2x Stacking Buffer Recipe (makes 1000ml)  
Tris HCl (pH 6.8) 30.35 g
SDS 2.0 g
Dissolve compounds thoroughly. Adjust pH slowly to pH 6.8 with concentrated HCl,  then add ddH2O to 1000ml.
1x Running Buffer Recipe (makes 1000ml)  
Tris-base 1.51 g
Glycine 7.5 g
SDS 0.5 g
Dissolve compounds thoroughly, then add ddH₂O to 1000 ml.
Tricine Gel Recipe      
Reagents Stacking Intermediate Separating
Gel percentage 4% 10% 15%
Gel volume 2ml 3ml 6ml
38% Glycerol - - 1.6
ddH₂O 1.4 1.2 -
30% Acrylamide 0.3 0.8 2.7
3.0 M Tris HCl (pH 8.5)
 - 1 2.14
1.0 M Tris HCl (pH 6.8)
 0.3 - -
10% SDS 0.02 0.03 0.06
10% APS 0.02 0.03 0.06
TEMED 0.002 0.003 0.003

IHC实验相关试剂配方

Citrate buffer For 1000 ml 1x TBS For 1000 ml
10 mM Trisodium citrate+2H₂O
 2.9 g 20 mM Tris-base 2.4 g
1.9 mM Citric acid+H₂O
 0.4 g 150 mM NaCl 8.7 g
Adjust pH to 6.0   Adjust pH to 7.6  
Add ddH₂O to 1000 ml   Add ddH₂O to 1000 ml  
Tris-EDTA (TE) buffer  For 1000 ml  1x TBST For 1000 ml
 10 mM Tris-base  1.21 g  1x TBS  999 ml
1 mM EDTA C₁₀H₁₄N₂Na₂O₈ +2H₂O
  0.372 g  Tween-20  1 ml
Adjust pH to 9.0      
Add ddH₂O to 1000 ml

 

IF实验相关试剂配方

1X PBS In 1000 ml (final volume)
10 mM Na₂HPO₄ 1.42 g
1.8 mM NaH₂PO₄ 0.22 g
140 mM NaCl 8.18g
2.68mM KCL 0.20 g
Adjust to pH 7.4  
Add ddH₂O to 1000 ml  
Antibody dilution buffer In 20 ml (final volume)
1% BSA 0.2g
Add 1X PBS to 20ml  

IP实验相关试剂配方

RIPA lysis buffer For 1000 ml
50 mM TrisHCl, pH 7.4 (1 M stock) 50 ml
150 mM NaCl 8.76 g
1% Triton X-100 10 ml
0.5% Sodium Deoxylcholate 5 g
0.1 % SDS 1 g
10 mM NaF 0.41 g
1 mM EDTA (0.5 M stock) 2 ml
Add ddH₂O to 1000 ml  
Adjust to pH 7.4  
Add PMSF to 1 mM and other protease inhibitors immediately prior to use. 
5X SDS sample buffer  
250 mM Tris HCl (pH 7.0) (1M stock) 12.5 ml
35% Glycerol 17.5 m
10% SDS 5 g
0.02% Bromophenol Blue 10 mg
10% ß-mercaptoethanol 5.0 ml
Add ddH₂O to 50ml, aliquot and store at -20°C

 

ChIP实验相关试剂配方

FC实验相关试剂配方

Blocking Buffer 1000 ml
Bovine serum albumin 5.00 g
1x PBS buffer 1000 ml
PBS Buffer 1000 ml
10 mM Na₂HPO₄ 1.42 g
1.7 mM NaH₂PO₄ 0.20 g
140 mM NaCl 8.19 g
Add ddH₂O to 1000 ml
Adjust to pH 7.4

 

包涵体蛋白纯化实验相关试剂配方

TNMFX-2M Urea For 1000 ml
50 mM Tris-base 6.06 g
150 mM NaCl 8.77 g
1 mM EDTA 0.37 g
2 M Urea 120.20 g
Adjust to pH 8.0
Add ddH₂O to 1000 ml
PBST buffer
For 1000 ml
58 mM Na₂HPO₄ 8.24 g
17 mM NaH₂PO₄ 2.04 g
68 mM NaCl 3.98 g
1%Triton-X100 10 ml
Adjust to pH 7.4
Add ddH₂O to 1000 ml
TNMFX-0.1% Triton X100 For 1000 ml
50 mM Tris 6.06 g
150 mM NaCl 8.8 g
1 mM EDTA 0.4 g
0.1% Triton-X100 1 ml
Adjust to pH 8.0
Add ddH₂O to 1000 ml
PBS with 2% Sarkosyl For 200 ml
58 mM Na₂HPO₄ 1.65 g
17 mM Na₂HPO₄ 0.41 g
68 mM NaCl 0.80 g
2% Sarkosyl 4 .00 g
Adjust to pH 8.0
Add ddH₂O to 200 ml
8 M Urea For 200 ml
Urea 96.08 g
Adjust to pH 8.0
Add ddH₂O to 200 ml


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