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. 2018 Aug 2;(138):57996. doi: 10.3791/57996
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