[Histonet] IHC for p21, p27, and RB

[Elizabeth Chlipala]

Richard

We just started working on a IHC protocol for p21 we tried the antibody from Dako, we looked at 1:25 and 1:50 at one hour RT incubation with pH9 retreival on some colon CA blocks, stain is specific to the nuclei with no background so far, but that's about all we have done, we were going to try an overnight incubation just to see what happens, overall we are not seeing a large amount of nuclei stain.  It cross reacts with rat - that's the species we are working with.  The rat samples stained but only a few nuclei.  Both samples were formalin fixed.

For p27 we have run this antibody just a couple times and it seems to work fine in our hands with FFPE tissues, we use human tonsil as a control.  We used the one from Santa Cruz since our target was also rat.  It worked well in both the human tonsil sample and the rat ovarian tumor we stained.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com

Ship to address:

1567 Skyway Drive, Unit E
Longmont, CO 80504

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Richard Cartun
Sent: Friday, May 25, 2012 11:25 AM
To: Histonet
Subject: [Histonet] IHC for p21, p27, and RB

What has been people's experience with immunohistochemical staining for p21, p27, and Retinoblastoma (RB) gene protein on formalin-fixed, paraffin-embedded human tissue?  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596 Office
(860) 545-2204 Fax



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