David, The only insight I might have is a paper describing non-specific binding of Cascasde Blue to murine bone marrow cells (Cytometry 37:60-67, 1999); we've reproduced this data and it does seem to be B lineage specific. Are your CD19+ cells the ones showing the non-specific staining? Kevin Kevin L. Holmes, Ph.D. Head, Flow Cytometry Section Research Technologies Branch NIAID, NIH Bldg. 7, Room 01 NIAID, NIH Phone: 301-496-9071 FAX: 301-402-4532 Email: kholmes@niaid.nih.gov -----Original Message----- From: David Dombkowski [mailto:helix.mgh.harvard.edu@helix.mgh.harvard.edu] Sent: Tuesday, November 07, 2000 12:47 PM To: cyto-inbox Subject: Cascade Blue I have stained PBL after ficol with CD3-biotin,CD19-biotin and CD16-biotin followed by Avidin-Cascade Blue from Molecular Probes. I have used about 1ug/sample of Avidin-Cascade Blue and have found a non-specific very bright stained population in all three samples. My impression is that Cascade Blue Dye is binding non-specifically for some reason. I am exciting dye with about 100MW of 407nm Kryptonn Laser line and collecting emission with a 450nm/65 Band Pass filter. If anyone has an idea what I am seeing with this bright non-specific staining I would appreciate comment. Thanks. David M. Dombkowski dombkowski@helix.mgh.harvard.edu Flow Cytometry-Pathology-CNY rm7017 149 13th Street Massachusetts General Hospital-East Boston, MA 02129 Tel. (617)726-1683 Fax.(617)724-3164
This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:39 EST