Hi Amanda,
Coomassie blue will not interfere with the digestion itself. You can remove some of it with some 20% methanol without losing the proteins (be careful: nitrocellulose will dissolve in
strong solvents). If your digests is analyzed by LC-MS, then the coomassie is of no concern. If you are thinking of MALDI-ToF, a Zip-Tip cleaning is best. From what I remember, you
get more problems from the blocking reagents (we used PVP-40 at the time) than from the Coomassie.
You can write me directly if you need more precise info on this.
Axel
Amanda Hall wrote:
> Hi everyone,
>
> Leading on from my question from last week - can you do a tryptic digest on nitrocellulose if the NC has been stained with Coomassie? Can you just remove the CB with acetonitrile?
>
> Thanks
>
> Amanda
>
> |\ _,,,---,,_
> zz /,`.-'`' -. ;-;;,_
> |,4- ) )-,_..;\ ( `'-'
> '---''(_/--' `-'\_)
>
> Amanda Hall-Griffin
> Professional Officer
> Newcastle Protein
> University of Newcastle
> Ph (02) 4921-7299
> Fax (02) 4921-6903
-- ------------------------------------------------------------------------Axel Ducret, Ph.D. Senior Research Biologist Merck-Frosst Canada Inc. Dept. Biochemistry and Molecular Biology P.O. Box 1005 Pointe-Claire-Dorval PQ H9R 4P8 Canada
tel. + (514) 428-3428 fax + (514) 428-4900
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