[Histonet] (no subject)

[Vijayalakshmi Ananthanarayanan]

Hi All,

I am a postdoc fellow and I work a lot with formalin fixed paraffin embedded
human prostate specimens.
I am currently working on a DNA oxidation antibody (8-OHdG -mouse
monoclonal) from QED. My problem is that I get an awful lot of background,
which does not disappear with antibody dilutions (this also considerably
weakens the true positives). I use PBS for my washes and the primary is
incubated overnight at RT (as per the company protocols) and the secondary
is the Envision + system from DAKO. I use the ready to use peroxidase and
protein block from DAKO. My negative controls are fine and there is no
staining in them.
Also, I do not have a suitable positive control...
I would like to ask the following questions:
a) Has anyone tried using this antibody?
b) Does anyone know of a suitable positive control?
c) How do I eliminate the background? Is it better to use Tris buffer with
Tween rather than PBS?

Thanks,
Viju.

Dr.Vijayalakshmi Ananthanarayanan M.D.
Feinberg School of Medicine,
Northwestern University
Department of Preventive Medicine
680 N. Lake Shore Drive, Suite 1102
Chicago, IL  USA 60611
viju <@t> northwestern.edu
Phone:312-503-3122.
Pager:5-3269.
Fax: 312-908-9588