Hi!

I'd like to try the protocol described by Mineki et al., but I'm confused about the ratio of acrylamide : protein concentration in order to achieve proper alkylation of my free thiols and limit alkylation to other residues.

Mineki R, Taka H, Fujimura T, Kikkawa M, Shindo N, Murayama K. In situ alkylation for identification of cysteinyl residues in proteins during one- and two-dimensional sodium dodecyl sulphate–polyacrylamide gel electrophoresis. Proteomics 2002;2:1672–1681

Thanks in advance!

Mariana