quantification of protein
AFS7 at le.ac.uk
Wed Sep 22 17:26:55 EST 1999
Richard P. Grant wrote:
> To double check your total protein loading (it's conceivable that if the
> populations of inhibited and non-i are very different your quant method
> could be affected) you'd probably want to detect a 'house-keeping' protein
> on the same blot: this would check that you weren't lowering protein
> expression across the board, i.e. make you happier yopu're seeing a
> specific effect.
There's always the risk that your housekeeping protein could also be
affected by the antibiotic.
Alternative (or as well) you could Ponceau-S stain the membrane after
the blot. I would _always_ do this for westerns in any case, because
it's so quick and can save you from wasting time and reagents if the
blot or gel hasn't worked properly. Coomassie staining the gel after
blotting to see how much protein was left behind is also a good idea.
More information about the Cellbiol