Roger,
The following protocol is from "Current Protocols in Protein Science",
1. In a microcentrifuge tube, dissolve protein in a minimal volume (<100
ul if possible) of 0.3 M Tris-HCl pH 8.3, to give a protein concentration
>100ug/ml
2. If reduction is desired, add DTT to yield 10 fold molar excess
relative to expected disulfides
3. Add acrylamide to final concentration of 2 M. Incubate 2 hr at 37 C.
4. Either quench the reaction with 2.5 ul of 10% betamercaptoethanol and
desalt, or desalt immediately with a rapid technique such as RP-HPLC or
membrane absorption using a ProSorb cartridge,
Jacek Mozdzanowski
Analytical Sciences
SmithKline Beecham