Solution recipe for 500mL Calmodulin-Binding Buffer.
| 1. Measure 12.5mL of 1M Tris pH 8.0 into 100mL millipore H2O in a 500mL beaker
| 25mM Tris-HCl
|
| 2. Add 15mL of 5M NaCl
| 150mM NaCl
|
| 3. Add 500 μL of 1M Magnesium Acetate
| 1mM MgOAc
|
| 4. Add 500 μL of 1M Imidazole
| 1mM Imidazole
|
| 5. Add 1 mL 1M CaCl2
| 2mM CaCl2
|
| 5. Bring volume to 500mL with millipore H2O in a graduated cylinder
| Volume to 500mL
|
| 6. Complete immediately prior to use with 0.696 μL β-mercaptoethanol per 1 mL CBB
| 10mM β-mercaptoethanol
|
| 7. Keep completed CBB on ice at all times
|
