Modified CSF-XB (MCSF-XB)
1× XB salts, dilute from Extract buffer (XB) salts (20×)
1 mm MgCl2
50 mm sucrose (e.g., Sigma-Aldrich SX1075; dilute from a 1.5 m solution that has been filter-sterilized and stored at −20°C)
10 mm HEPES (e.g., Sigma-Aldrich H3375; dilute from a 1 m, pH 7.7 solution that has been filter-sterilized and stored at −20°C)
5 mm ethylene glycol tetraacetic acid (EGTA; e.g., Sigma-Aldrich 324626; dilute from a 0.1 m solution)
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Freshly prepare 250 mL (or more as needed). Adjust pH to 7.7 using KOH.
Just before use, add 1:1000 of each of:
Chymostatin (e.g., Roche 11004638001; 10 mg/mL in ethanol that has been aliquoted and stored at −20°C)
Leupeptin (e.g., Sigma-Aldrich 11017128001; 10 mg/mL in DMSO that has been stored at −20°C)
Pepstatin A (e.g., Sigma-Aldrich 516485; 10 mg/mL in DMSO that has been aliquoted and stored at −20°C)
