I do not know of a non mouse anti human macrophage marker, but in my experience most are using the wrong ms monoclonal anti human CD68 for macrophage markers, The KP1 clone will stain a lot more than macrophages, granulocytes etc., the best clone to use for anti human CD68 is the PGM1 clone which is specific to just macrophages and monocytes and not all the other cells. I have known hematopathologist who tell me they stopped using CD68 because it was just not specific enough, but when I tell them to switch to the PGM1 clone they are very happy with that.
As for macrophages in mouse tissue, I use the serotec rat anti mouse F4/80 ab with very good results. I have used macrophage markers that are for rat tissue (can't remember what they are at the moment, I think the are CD68?) but I did not have as good results on rat tissue with them as I did on ms tissue with rat anti f4/80. Liz, What do you use for macs on rat tissue? I may have asked you about this before? Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Isaac Milos Sent: Friday, October 23, 2009 1:57 PM To: Liz Chlipala; Margaryan, Naira; Galbraith, Joe; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] human macrophages in human melanoma xenografts Liz, Great point, and definitely something to keep in mind should one do that stain. Indeed, CD14 is not specific to macrophages only (monocytes also express the antigen as discussed). Of course, there are other more specific macrophage markers (i.e. HAM56) but these are commonly developed in mice. The CD14 came to mind as ours is a rabbit antibody. Best regards, Isaac Milos Cell Marque Corporation -----Original Message----- From: Liz Chlipala [mailto:l...@premierlab.com] Sent: Friday, October 23, 2009 11:15 AM To: Isaac Milos; Margaryan, Naira; Galbraith, Joe; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] human macrophages in human melanoma xenografts She is asking to stain for macrophages and that's a monocyte/macrophage marker so that's not the same. You have to be careful with the names of antibodies and check the specification sheets. You can search for a macrophage marker and the antibody may be listed as a macrophage marker as the following example: This antibody is from abcam its listed as follows: Macrophage antibody [MAC387] (ab22506) but when you look at the specification sheet it states -- Recognises the L1 or Calprotectin molecule, an intracytoplasmic antigen comprising of a 12kD alpha chain and a 14kD beta chain expressed by granulocytes, monocytes and by tissue macrophages. So it is not entirely specific to macrophages. This antibody may work fine for your application, but if you are looking just to stain macrophages this is not the antibody you want to pick. Just a word of advice. 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: Isaac Milos [mailto:imi...@cellmarque.com] Sent: Friday, October 23, 2009 12:01 PM To: Margaryan, Naira; Liz Chlipala; Galbraith, Joe; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] human macrophages in human melanoma xenografts Hi Naira, Our new IVD rabbit monoclonal (anti human) CD14 is preferentially expressed on monocytes/macrophages: http://www.cellmarque.com/07/p_detail.php?id=2066 Best regards, Isaac Milos Cell Marque Corp. -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Margaryan, Naira Sent: Friday, October 23, 2009 8:57 AM To: Liz Chlipala; Galbraith, Joe; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] human macrophages in human melanoma xenografts Thanks a lot both of you! For melanoma detection, I usually use HRP with AEC. Is there any non-mouse anti-human macrophage marker? Thank you much, Naira -----Original Message----- From: Liz Chlipala [mailto:l...@premierlab.com] Sent: Friday, October 23, 2009 10:51 AM To: Galbraith, Joe; Margaryan, Naira; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] human macrophages in human melanoma xenografts F4/80 is a mouse macrophage marker. If she wants to detect human macrophages in a mouse background she will need to use a mouse anti-human CD68. She will need to run it with a mouse on mouse detection system and run all of the appropriate negative controls. I would also select a alkaline phosphatase detection system rather than HRP with DAB just incase the tumor has any melanin in it. I have done this before looking for human lymphocytes with LCA in a mouse background. You just need to make sure you have all of the appropriate controls. For a positive control I would use a human tonsil or something like that and you need to make sure you run the appropriate isotype negative controls. 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 Galbraith, Joe Sent: Friday, October 23, 2009 9:44 AM To: Margaryan, Naira; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] human macrophages in human melanoma xenografts Naira: Here is a link to a site listing macrophage markers. F4/80 is a commonly mentioned marker for macrophages. I presume you mean IHC rather than ICH. Enjoy. http://www.antibodybeyond.com/reviews/cell-markers/macrophage-marker.htm Joe joseph-galbra...@uiowa.edu -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Margaryan, Naira Sent: Friday, October 23, 2009 10:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] human macrophages in human melanoma xenografts Hi Histonetters, I would like to be able to look at human macrophages in human melanoma xenografts raised in mouse. Could you please suggest me a best Ab for ICH and protocol? Thanks in advance, Naira _______________________________________________ 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 _______________________________________________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet