Solution Preparation
1. 1M DTT (dithiothreitol)
- 7.7mg DTT in 50uL H2O
or
- 38.5 mg in 250uL H20
2. Ellman's Reagent (0.4% 5,5'-dithiobis(2-nitrobenzoic acid))
- 4mg Ellman's Reagent in 1mL 100mM TEA pH 8
3. 3mM FMOC-NHS
- dissolve 0.5mg FMOC in 300uL DMSO; add 200uL 100mM TEA pH 7
4. MPS (3-mercaptopropyl silane)
- 40mL ethanol, 2 drops Glacial Acetic Acid, 0.3mL MPS
5. 1mM MUAM
- 0.9mg MUAM in approximately 7mL EtOH
6. 4mM PEG-NHS
- 2mg PEG in 250uL of 100mM TEA pH 8
7. 1mM SSMCC
- 0.4mg SSMCC in 1mL of 100mM TEA pH 7
8. 1M TAEA (tris(2-aminoethyl)amine)
9. 100 mM TEA pH 7 & 8
10. 1M TEAA pH 7.01 (triethylammonium acetate)
11. Phosphate buffer
| Component | Stock Solution | 20
mL | 250 mL |
| 20mM phosphate | 200 mM pH 7.74 | 2 mL | 25 mL |
| 100mM NaCI | 2 M | 1 mL | 12.5 mL |
| 1 mM EDTA | 0.1 M | 200 uL | 2.500 mL |
| 1 mM DTT | 1 M | 20 uL | 250 uL |
| 5mM MgCl2 | 1 M | 100 uL | 1.25 mL |
| Final Dilution | Milli-Q | 20 mL | 250 mL |
Summer 1999
JB, BPN, ed.
Copyright 1999 Corn Group