I have a client who's used Red-Out. I think it's a polyclonal anti Glyc A that aggregates the RBC's. Robbins Scientific Corp Sunnyvale CA USA 800-752-858 or 408-734-5800 Dennis ************************************************************************* * Dennis J. Young Voice : (619) 822-0407 * * Flow Cytometry Core Facility FAX : (619) 822-0412 * * University of California, San Diego USA e-mail: djyoung@ucsd.edu * * http://www-core.ucsd.edu/flow-core.html * ************************************************************************* ______________________________ Reply Separator _________________________________ Subject: Cord blood Author: williams@ichr.uwa.edu.au at @UCSD Date: 7/31/97 1:10 PM Hi Flow people I'm working on cord blood phenotyping, interested in rare leukocyte subsets. As those who work on these preps will know, there is a problem with RBC and erythroid contamination, variable but up to 80% of ficoll separated PBMC. They don't seem to lyse easily using hypotonic methods, and as some are nucleated, I wouldn't expect this to be an ideal approach. Gating on FS/SS can partly improve percentage of CD45+ cells but not much. We really want to clean the preps up, and I have been thinking of using complement lysis of the RBC, perhaps with an antibody to Glycophorin A. There are antibodies of different isotypes available, including IgM, I presume this might be the best to start with for complement lysis. I believe GlyA has a complement regulatory function, I don't know if this will be a problem. Does anybody have any hints on this, either the lysis method, or is there a simpler way of removing the erythroid cells that I have missed out on? Immunomagnetic separations can get a bit expensive, and positive selection of CD45+ cells will cause problems with further analysis. I presume there must be a lot of people doing this sort of thing, because of the CD34 interest. Thanks. ______________________________________________ Dr William Smith Institute for Child Health Research (Company Limited by Guarantee ACN 009 278 755) Subiaco, Western Australia, 6008 PO Box 855 West Perth WA 6872 Ph 61 8 9340 8792/8388, Fax 61 8 9388 3414 email williams@ichr.uwa.edu.au ______________________________________________ >-- Saved internet headers (useful for debugging) >Received: from flowcyt.cyto.purdue.edu by mail.ucsd.edu; id NAA18196 sendmail 8 >Received: by flowcyt.cyto.purdue.edu (940816.SGI.8.6.9/930416.SGI.AUTO) for cyt >Received: from ichr.uwa.edu.au by flowcyt.cyto.purdue.edu via ESMTP (940816.SGI >References: TVWTICHR, Company Limited by Guarantee, ACN 009 278 755 >Received: (from uucp@localhost) by ichr.uwa.edu.au (8.8.5/8.8.5) id NAA29405 fo >Received: from cbi-mac-wills.ichr.uwa.edu.au(130.95.224.198) by ichr.uwa.edu.au >Message-Id: <l03102801b005cc9aa12d@[130.95.224.198]> >Mime-Version: 1.0 >Content-Type: text/plain; charset="us-ascii" >Date: Thu, 31 Jul 1997 13:10:53 +0800 >To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >From: Dr William Smith <williams@ichr.uwa.edu.au> >Subject: Cord blood
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:58 EST