Mark I took a brief look at the abcam antibody and in particular the review with the paraffin staining. If I was purchasing the antibody I would question the results of that review, since they used a goat anti-mouse detection system which is not ideal for mouse tissue. Plus I'm not sure that the staining pattern is consistent with what I would expect. I would expect to see staining of the myofibrils within the cell. It appears from the image to be staining nuclei and the outside of the cell. I would call Abcam to see if they have tested this antibody internally for paraffin processing or did they completely rely on that review. You could also try enzyme retrieval rather than HIER.
Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Elliott Sent: Wednesday, March 02, 2011 3:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mouse cardiac myosin antibodies I have been asked to stain mouse hearts for cardiac myosin. I have tried the antibody from Abcam (ab50967-Mouse monoclonal to heavy chain cardiac myosin) with no luck. I am using the Mouse on Mouse kit from Biocare and it worked well for me for some other mouse antibodies on mouse tissue but doesn't want to work on this antibody. I have tried different retrieval solutions (pH6 and pH9, as well as Biocare rodent Decloaker). Anyone use this antibody? Any suggestions for another antibody greatly appreciated. Thanks Mark ***CONFIDENTIALITY NOTICE*** This electronic message and any attachments are intended only for the use of the addressee and may contain information that is privileged and confidential. Any dissemination, distribution or copying of this communication by unauthorized individuals is strictly prohibited. If you have received this communication in error, please notify the sender immediately by reply e-mail and delete the original and all copies from your system. _______________________________________________ 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