Trouble wt.S-Tag western blot kit

Max Soegaard tmms at mbio.aau.dk
Wed Aug 19 10:49:51 EST 1998


Dear netters.

I have tried to use Novagen's S-Tag western blot kit to detect a protein
that is fused to the S-peptide. This protein is secreted out of my AMA-
cells and I want to be able to assay for its prescense in the medium by
just sampling the medium, run it on a PAA- gel and then do the western
blot.  My problem is specificity. The HRP-S-protein seems to bind to
just about everything from the cell medium (DMEM + 10% NCS) so my
western blots look pretty similar to comassie-stains - and I dont see
anything specific...

Has anyone experienced something at all like this with the S-tag western
blot kit from Novagen? What did you guys do about it? Has anybody had a
good experience with this western blot kit?
Any input (at all) will be greatly appriciated..

Yours
Max Soegaard
Inst. of Struct. and Mol. Biol.
University of Aarhus.
Denmark.
email: tmms at mbio.aau.dk




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