Yu:TEV Cleavage Buffer

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[[Category:Yu Solutions]]

Revision as of 17:25, 22 September 2009

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Solution recipe for 500mL TEV Cleavage 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. Immediately prior to use, complete with: Complete with:
Protease inhibitors for completing CBB
1μL of fresh 1M DTT per 1mL of buffer 1mM DTT
1μL of 1M Imidazole per 1mL of buffer 1mM Imidazole
1μL 1M magnesium acetate per 1mL of buffer 1mM MgOAc
7. Keep completed CBB on ice at all times
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