Yu:TEV Cleavage Buffer: Difference between revisions
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Michaelcyu (talk | contribs) (TEV cleavage buffer) |
Michaelcyu (talk | contribs) (CBB) |
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|Volume to 500mL | |Volume to 500mL | ||
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|6. Immediately prior to use, complete with 1μL of fresh 1M DTT per 1mL of buffer | |6. Immediately prior to use, complete with: | ||
| | |Complete with: | ||
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|+ '''Protease inhibitors for completing CBB''' | |||
|1μL of fresh 1M DTT per 1mL of buffer | |||
|1mM DTT | |||
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|1μL of 1M Imidazole per 1mL of buffer | |||
|1mM Imidazole | |||
|- | |||
|1μL 1M magnesium acetate per 1mL of buffer | |||
|1mM MgOAc | |||
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|7. Keep completed CBB on ice at all times | |||
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|} | |} |
Revision as of 12:15, 14 June 2009
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. Immediately prior to use, complete with: | Complete with: | ||||||
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7. Keep completed CBB on ice at all times |