Check out Azpiroz, R. and R. Butow (1995), Methods Enzymol. 260, p.? There
is a simple method to preclear antibodies with protein extracts adsorbed
to nitrocellulose. It works like a charm, even on really dirty antisera.
Affinity jpurification is nice, but it is laborious and kills the antibody
within weeks after purification.
Specifically,
Azpiroz, R.
On 4 Oct 1996 GMEACHAM at bmg.bhs.uab.edu wrote:
To all,
I am doing indirect immunofluorescence on S.cerevisiae YPH499. I am
having a problem with high backround i.e. the preimmune control lights
up like a Christmas tree. We are about to affinity purify the Ab. What
fixatives or levels of blocking has anyone had success in reducing
backround? Our Ab looks pretty good on a western. Thanks in advance.
Geoff Meacham
UAB Dept. of Cell Biology
----------------------
Ricardo Azpiroz
Department of Biochemistry
Life Sciences South
University of Arizona
Tucson AZ 85721-0106
azpiroz at U.Arizona.EDU
