Amos u might have to get an anti rabbit link that has been mouse absorbed. Dako's anti rab envision is made in a goat but it is not mouse absorbed. Southern Biotec or Jackson Labs are really good places to get absorbed links. Still no reliable cd4 and cd8 for ffpe mouse tissue as far as I know, I have tried many that claim they work but have not consistently worked in my hands, we still do that with frozen mouse tissue not aldehyde fixed. Are u doing HIER with the EDTA ph9? In my experience especially with the higher ph u can over retrieve and cause undesirable nuclear staining.
Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting 40864 E. Arkansas Ave Bennett, CO 80102 H 303-644-4538 C 720-281-5406 prueg...@hotmail.com rueggihcconsultin...@outlook.com This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: Friday, January 24, 2014 10:09 AM To: histonet@lists.utsouthwestern.edu; ihcrg Group, (E-mail) Subject: [Histonet] Mouse GranzymeB Ok Musketeers, I am trying to detect cytotoxic T-Cells in formalin fixed paraffin embedded mouse liver. GranzymeB should do the trick. I have a rabbit anti mouse (and human and rat) GranzymeB from abCam (ab53097). The spleen that I ran as a control with it works fine. Nice T-cells and no signifigant background staining. The liver on the others hand has some lymphocyte staining, presumably cytotoxic T-cells as expected, but it looks like every hepatocyte nuclei are also picking it up. Grrr, right? If this antibody had been raised in mouse, I wouldn't be surprised. Also, if the spleen looked similar, I wouldn't be surprised. Do any of you have any ideas ab pi ut what this might be? Alternatively, does anyone have any ideas about another antibody that would work here? CD4 or CD8 would be great here, but its FFPE mouse, so sadly that's out (more grumbling). For the completionists out there, I used EDTA pH9, peroxide block and detected the antibody (diluted 1:800) with Envision (rabbit) and DAB from Dako. Any suggestions would be vastly appreciated, Happy Friday, Amos _______________________________________________ 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