Calmodulin binding buffer 1 (CBB1)

Reagent	Final concentration	Amount
Tris-HCl (pH 8.0) (1 M)	10 mM	0.5 mL
NaCl (5 M)	150 mM	1.5 mL
Magnesium acetate (pH 7.4) (1 M)	1 mM	0.05 mL
Imidazole (pH 9.0) (1 M)	1 mM	0.05 mL
CaCl2 (1 M)	2 mM	0.1 mL
Glycerol (50%)	10%	10 mL
2-Mercaptoethanol	10 mM	0.035 mL
NP-40 (10%)	0.1%	0.5 mL
ddH2O	n/a	37.265 mL
Total	n/a	50 mL

Prepare on the day of the experiment, filter sterilize and keep at 4°C. Adjusting the pH after mixing all the ingredients is not needed.