. 2001 Feb 15;29(4):e24. doi: 10.1093/nar/29.4.e24

Table 3. Buffers.

Cell lysis buffers	1^a	50 mM Tris–HCl pH 8.0, 150 mM NaCl, 0.1% Triton X-100, 10 mM β-mercaptoethanol, 1 mM magnesium acetate, 1 mM imidazole, 2 mM calcium chloride, 1 mM DTT, 0.05% deoxycholic acid, 10 mM sodium fluoride and PIs^b
	2^a	50 mM NaH₂PO₄ pH 8.0, 150 mM NaCl, 10 mM imidazole, 50 mM sodium fluoride and PIs^b
Elution buffers	1^a	50 mM Tris–HCl pH 8.0, 250 mM NaCl, 50 mM sodium fluoride, 2 mM EGTA, 10 mM β-mercaptoethanol and PIs^b
	2^a	50 mM NaH₂PO₄ pH 8.0, 300 mM NaCl, 50 mM sodium fluoride, 250 mM imidazole and PIs^b
Immunoprecipitation buffer		50 mM Tris–HCl pH 7.5, 250 mM NaCl, 0.5% Triton X-100, 50 mM sodium fluoride, 5 mM EDTA and PIs^b
2× kinase reaction buffer		100 mM Tris–HCl pH 7.5, 20 mM MgCl₂, 2 mM DTT

^a1: used for calmodulin–peptide binding affinity column; 2: used for Ni affinity column.

^bPIs, protease inhibitors: 1.5 mM PMSF, 10 µg/ml leupeptin, 5 µg/ml pepstatin A, 10 µg/ml TPCK, 2 mM benzamidine, 0.2 mg/ml bacitracin.