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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet