Buffer compositions
| Application . | Buffer . | Composition . |
|---|---|---|
| ChIP | IP buffer | 150mM NaCl, 50mM Tris, 5mM EDTA, 0.5% NP-40, 1% Triton X-100, and 1× protease inhibitor cocktail from Roche |
| ChIP | Sonication buffer | 140mM NaCl, 50mM HEPES, 1mM EDTA, 1% Triton X-100, 0.1% NaDOC, 0.1% SDS, and 1× protease inhibitor cocktail |
| ChIP | Wash buffer I | 1% Triton, 0.1% NaDOC, 150mM NaCl, and 10mM Tris-Cl, pH 8 |
| ChIP | Wash buffer II | 1% NP-40, 1% NaDOC, 150mM NaCl, and 10mM Tris-Cl |
| ChIP | Wash buffer III | 0.5% Triton, 0.1% NaDOC, 500mM NaCl, and 10mM Tris-Cl |
| ChIP | Wash buffer IV | 0.5% NP-40, 0.5% NaDOC, 250mM LiCl, 20mM Tris-Cl, and 1mM EDTA |
| ChIP | Wash buffer V | 0.1% NP-40, 150mM NaCl, 20mM Tris-Cl, and 1mM EDTA |
| ChIP | TE | 10mM Tris-Cl, pH 8, and 1mM EDTA |
| Western blotting | Lysis buffer | 150mM NaCl, 10mM Tris HCl, 5mM EDTA, 1% NP-40, 0.1% SDS, 0.5% DOC, 1mM Na3VO4, 1mM NaF, 5mM Na pyrophosphate, and a protease inhibitor cocktail |
| Western blotting | TBST | 10mM Tris, pH 7.5, 154mM NaCl, and 0.15% Tween 20 |
| Application . | Buffer . | Composition . |
|---|---|---|
| ChIP | IP buffer | 150mM NaCl, 50mM Tris, 5mM EDTA, 0.5% NP-40, 1% Triton X-100, and 1× protease inhibitor cocktail from Roche |
| ChIP | Sonication buffer | 140mM NaCl, 50mM HEPES, 1mM EDTA, 1% Triton X-100, 0.1% NaDOC, 0.1% SDS, and 1× protease inhibitor cocktail |
| ChIP | Wash buffer I | 1% Triton, 0.1% NaDOC, 150mM NaCl, and 10mM Tris-Cl, pH 8 |
| ChIP | Wash buffer II | 1% NP-40, 1% NaDOC, 150mM NaCl, and 10mM Tris-Cl |
| ChIP | Wash buffer III | 0.5% Triton, 0.1% NaDOC, 500mM NaCl, and 10mM Tris-Cl |
| ChIP | Wash buffer IV | 0.5% NP-40, 0.5% NaDOC, 250mM LiCl, 20mM Tris-Cl, and 1mM EDTA |
| ChIP | Wash buffer V | 0.1% NP-40, 150mM NaCl, 20mM Tris-Cl, and 1mM EDTA |
| ChIP | TE | 10mM Tris-Cl, pH 8, and 1mM EDTA |
| Western blotting | Lysis buffer | 150mM NaCl, 10mM Tris HCl, 5mM EDTA, 1% NP-40, 0.1% SDS, 0.5% DOC, 1mM Na3VO4, 1mM NaF, 5mM Na pyrophosphate, and a protease inhibitor cocktail |
| Western blotting | TBST | 10mM Tris, pH 7.5, 154mM NaCl, and 0.15% Tween 20 |
DOC indicates deoxycholate.