Re: Immunostain for PCNA

Did ytou perfprm any pretreatments, specifically HIER?  We use HIER on our PCNA's.


02/27/03 03:14 PM

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        Subject:        Immunostain for PCNA

Hi. I was wondering if anyone has any suggestions on a protocol for PCNA stain. I'm staining mouse lung tissue using the Envision+ kit from DAKO. I'm using a rabbit anti-PCNA from Santa Cruz biotech as the primary Ab. I incubate at RT for 1hr. Everything else is done as suggested by the kit (peroxidase block, serum block, secondary Ab, chromogen, etc). I used the PCNA Ab at 1:50 and got nothing. When I went to 1:25, I had a lot of backrgound and no stain in the nucleus whatsoever. I'm wondering if anyone has any suggestions on this stain, and also what is the best tissue from mouse to use as a positive control. 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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