Hi Fabrice, If you are looking for NFkB activation I suggest to use antibody against non-phosphorylated p65 (rabbit from Abcam, CHIP grade) and watch nuclear translocation of p65. Different anti-phospho-p65 did not give me anything, while anti-p65 clearly shows NFkB activation every time (and it correlates very well with biochemical data). The best results I got was with fresh-frozen sections (with subsequent formaldehyde fixation), next - cryo sections from formaldehyde perfused mice or rats, next - cryo from formaldehyde fixed tissues, and finally - sometimes got some positive results from FFPE after retrieval by citrate boiling.
Anatoli Gleiberman, PhD Director of Histopathology Cleveland Biolabs, Inc 73 High Street Buffalo, NY 14203 phone:716-849-6810 ext.354 fax:716-849-6817 e-mail: agleiber...@cbiolabs.com -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Fabrice GANKAM Sent: Monday, September 20, 2010 6:50 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] multi-timer Hey guys Was wondering if any of you ever used a phopho NFKB p65 antibodu and which one you prefer with what antigen retrieval method if applies. I'm working on FFPE rat tissue Thanks. Fabrice _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This communication may contain privileged information. It is intended solely for the use of the addressee. If you are not the intended recipient, you are strictly prohibited from disclosing, copying, distributing or using any of this information. If you received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy. This communication may contain nonpublic information about individuals and businesses subject to the restrictions of the Gramm-Leach-Bliley Act. You may not directly or indirectly reuse or redisclose such information for any purpose other than to provide the services for which you are receiving the information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
