RE: MMP-2 and MMP-9 Immunostaining
We have been using the MMP-9 polyclonal antibody from Biogenesis Inc.
catalog #5980-0911 on FFPE sections. We antigen retrieve in citrate pH6,
standard pressure cooker method of 20 + 20, use ab dilution of 1:100 O/N at
4C. Secondary is 1:200, use HRP Strpetavidin detection at 1:250 and then
Chromagen of choice.
All that said, the antibody is somewhat variable, and most of our PI's have
moved away from ordering it.
Best of luck!
Center for Cardiovascular Research
University of Rochester Medical Center
KMRB Room 2-9816
601 Elmwood Avenue.
PO Box 679
Rochester, New York 14642
> From: NIDAL E MUVARAK
> Sent: Tuesday, March 4, 2003 7:07 PM
> To: HistoNet Server
> Subject: MMP-2 and MMP-9 Immunostaining
> Hi everyone. I've been performing MMP-2 and -9 on mouse lung tissue and
> pulmonary arteries. I got the primary antibodies from Santa Cruz Biotech,
> and I regret doing so since the staining is so variable. I'm using the
> Envision+ DAKO kit, and it's for rabbit primaries. I'm positive I'm doing
> everything correctly. I just can't figure out the right dilutions for the
> primaries. For every experiment, I get different staining, even when using
> the ab at the same dilution. So I was wondering if anyone suggests good
> primary Ab for MMP-2 and -9. Thank you for your time.
> Nidal E Muvarak
> Associate Research Specialist
> Department of Biomedical Engineering
> University of Wisconsin-Madison
> 1550 Engineering Dr.; Rm. 2158
> Madison, WI 53706-1609
> Lab: (608) 265-8921; Office: (608) 265-4205;
> Home: (608) 256-7934; Cell: (608) 332-6068
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