Does anyone have a suggestion for a method to florescently tag some dyes to the cell lysate proteins? I am searching for a bioconjugation method that can label most of the proteins in the lysate. Since this is a highly heterogeneous mixture of proteins, I am not sure if the conventional NHS ester reactions will be successful or whether this is a reasonable method to pursue. I'd appreciate it if some chemist/ biologist share their insight.
I think it will work. Almost all proteins contain accessible amino groups for labeling (lysine and the N-terminus). You will have to add a lot of the NHS ester, though, because there will be so many groups to label. Also, you will have to use a lysis buffer that does not contain any amino groups (e.g. no Tris buffer or HEPES).
Thanks. I tried the strategy with fluorophores with both NHS ester and maleimide dye mixtures and it seems to be working fine. Thanks again.
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