Buffer
|
Volume
|
Composition
|
6 M guanidinium chloride lysis buffer |
50 mL |
6 M guanidinium chloride, 100 mM tris pH 8.5, 10 mM tris (2-carboxyethyl) phosphine, 40 mM chloroacetamide, 2 mM sodium orthovanadate, 2.5 mM sodium pyrophosphate, 1 mM β-glycerophosphate, 500 mg n-octyl-glycoside, ultra-pure water to volume |
100 mM sodium pyrophosphate |
50 mL |
2.23 g sodium pyrophosphate decahydrate, ultra-pure water to volume |
1M β-glycerophosphate |
50 mL |
15.31 g β-glycerophosphate, ultra-pure water to volume |
5% trifluoroacetic acid |
20 mL |
Add 1 mL of 100% trifluoroacetic acid into 19 mL ultra-pure water |
0.1% trifluoroacetic acid |
250 mL |
Add 5 mL 5% trifluoroacetic acid to 245 mL ultra-pure water |
pY elution buffer |
250 mL |
0.1% trifluoroacetic acid, 40% acetonitrile, ultra-pure water to volume |
pST elution buffer |
250 mL |
0.1% trifluoroacetic acid, 50% acetonitrile, ultra-pure water to volume |
IP binding buffer |
200 mL |
50 mM tris pH 7.4, 50 mM sodium chloride, ultra-pure water to volume |
25 mM ammonium bicarbonate, pH 7.5 |
10 mL |
Dissolve 19.7 mg into 10 mL sterile ultra-pure water, pH to 7.5 with 1 N hydrochloric acid (~10-15 µL/10 ml solution), make fresh |
1M phosphate buffer, pH 7 |
1,000 mL |
423 mL 1 M sodium dihydrogen phosphate, 577 mL 1 M sodium hydrogen phosphate |
Equilibration buffer |
14 mL |
6.3 mL acetonitrile, 280 µL 5% trifluoroacetic acid, 1740 µL lactic acid, 5.68 mL ultra-pure water |
Rinsing buffer |
20 mL |
9 mL acetonitrile, 400 µL 5% trifluoroacetic acid, 10.6 mL ultra-pure water |
Mass spectrometry solution |
10 mL |
500 µL acetonitrile, 200 µL 5% trifluoroacetic acid, 9.3 mL ultra-pure water |
Buffer A |
250 mL |
5 mM monopotassium phosphate (pH 2.65), 30% acetonitrile, 5 mM potassium chloride,ultra-pure water to volume |
Buffer B |
250 mL |
5 mM monopotassium phosphate (pH 2.65), 30% acetonitrile, 350 mM potassium chloride, ultra-pure water to volume |
0.9% ammonium hydroxide |
10 mL |
300 μL 29.42% ammonium hydroxide, 9.7 mL ultra-pure water |