Recipe

Buffer H (elution, lysis, and wash)

Reagent Final concentration
Elution buffer H Lysis buffer H Wash buffer H
Glycerol 15% (v/v) 15% (v/v) 15% (v/v)
Imidazole 480 mM 20 mM 20 mM
NaCl 100 mM 500 mM 100 mM
Nonidet P-40 0.01% (v/v) 0.01% (v/v) 0.01% (v/v)
Sodium phosphate (pH 7.6) 50 mM 50 mM 50 mM
Store solutions for up to 24 h at 4°C.
Immediately before use, add:
β-glycerophosphate 10 mM 10 mM 10 mM
Benzamidine 0.5 mM 0.5 mM 0.5 mM
Phenylmethylsulfonyl fluoride 0.2 mM 0.2 mM 0.2 mM

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  1. doi:10.1101/pdb.rec11765 Cold Spring Harb Protoc 2009: pdb.rec11765-
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