n terminal deblocking of protein in gel or pvdf membrane
houthaeve at embl-heidelberg.de
Fri May 19 02:50:43 EST 1995
<barbara.valsasina at itner.pharmacia.se> wrote:
> sometimes I have to deal with proteins electroblotted onto pvdf
> membrane or in sds page that are impossible to sequence because they
> are blocked at the N terminal end .
> I usually try CNBr cleavage but after that I have problems in the
> extraction of obtained peptides.
> I would like to know if anybody has any experience in deblocking
> blotted proteins as for example with methanolysis or in extraction
> from pvdf (I use Problott membranes - from Applied Biosystems).
Just forget it. You have, when having not enough material, hardly any
chance to deblock !
What do you exactly want ?
Straightforward, I'd say, perform a peptide map with trypsin and check
on MALDI. Then fish out the peptide + eventually the blocking group.
More information about the Proteins