Yu:CBB: Difference between revisions
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|+'''Solution recipe for 500mL Calmodulin-Binding Buffer.''' | |+'''Solution recipe for 500mL Calmodulin-Binding Buffer.''' | ||
|width="600px" valign="top"|1. Measure 12.5mL of 1M Tris pH 8.0 into a 500mL beaker | |width="600px" valign="top"|1. Measure 12.5mL of 1M Tris pH 8.0 into 100mL millipore H<sub>2</sub>O in a 500mL beaker | ||
|width=" | |width="150px" valign="top"|25mM Tris-HCl | ||
|- | |- | ||
|2. Add 15mL of 5M | |2. Add 15mL of 5M NaCl | ||
| | |150mM NaCl | ||
|- | |- | ||
|3. Add | |3. Add 500 μL of 1M Magnesium Acetate | ||
| | |1mM MgOAc | ||
|- | |||
|4. Add 500 μL of 1M Imidazole | |||
|1mM Imidazole | |||
|- | |- | ||
| | |5. Add 1 mL 1M CaCl<sub>2</sub> | ||
| | |2mM CaCl<sub>2</sub> | ||
|- | |- | ||
|5. Bring volume to 500mL with millipore H<sub>2</sub>O in a graduated cylinder | |5. Bring volume to 500mL with millipore H<sub>2</sub>O in a graduated cylinder | ||
|Volume to 500mL | |Volume to 500mL | ||
|- | |- | ||
|6. Complete | |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 | |7. Keep completed CBB on ice at all times | ||
|} | |} | ||
Latest revision as of 12:28, 22 December 2009
| 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 |
