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Solution recipe for 500mL Calmodulin-Binding Buffer.
1. Measure 12.5mL of 1M Tris pH 8.0 into a 500mL beaker for a final concentration of 25mM Tris pH 8.0 12.5mL 1M Tris pH 8.0
2. Add 15mL of 5M NaCl for a final concentration of 150mM NaCl 15mL 5M NaCl
3. Add 5mL of 10% NP-40 for a final concentration of 0.1% NP-40 5mL 10% NP-40
4. Add 500μL of 0.5M EDTA for a final concentration of 0.5mM EDTA 500μL 0.5M EDTA
5. Bring volume to 500mL with millipore H2O in a graduated cylinder Volume to 500mL
6. Complete with protease inhibitors immediately prior to use: Complete:
Protease Inhibitors for CBB
Add 1μL of DTT per 1mL CBB 1mM DTT
Add 1μL of Imidazole per 1mL CBB 1mM Imidazole
Add 1μL of magnesium acetate per 1mL CBB 1mM MgOAc
7. Keep completed CBB on ice at all times