I am trying to perform non-lysing colony immunoblots of yeast, in
order to detect secretion of CPY (carboxypeptidase Y).
I have done this several times before with great results. Now,
however, I am having a tremendous problem with background. In fact, a
ran a negative control - just a piece of nitrocellulose, without any
cells. I got a very high level of background.
I just ordered a new ECL detection kit from Amersham - the same
kit I have always used. This kit has the detection reagents and the
secondary antibody. The nitrocelluose was the same.
Does anyone have any ideas? It is frustrating that something I
have done so well previously suddenly turns sour. I would appreciate any
Graduate Student - Cell & Molecular Biology
CSU Long Beach.
lgeller at csulb.edu, demode101 at mail.earthlink.net