We have a Fellow in the lab who would like to measure immunoregulatory CD4 t cells as part of his project. These cellos are reported to constitutively express CD25. The literature shows them as a dimly positive cluster, although much of the work has used bead isolation and I'm wondering if that is modifying the receptor experssion. We have spent the last 2 months trying to figure out how to do this. We have performed whole blood labeling with the thinking that the less sample manipulation we do , the better. We have also looked at Ficolled PBMC. We have used two different direct label antibodies [same clone, different fluors one FITC and one APC ] . We have also tried biotin CD25 followed by streptavidin FITC and APC. Each method has yielded slightly different values. We tried the enzyme amplification system without success. We are now ordering a different clone of CD25 McAb from a different vendor. It's unclear to me what's the best way to quantify thsi population. Any insight on how to phenotypically identify this subpopulation would be appreciated. Thanks, Tom **************************************************************************** * Thomas W. Mc Closkey, Ph. D. Director of Flow Cytometry, North Shore University Hospital Assistant Professor of Pediatrics, New York University School of Medicine Boas Marks Biomedical Research Center, 350 Community Drive Manhasset, Long Island, New York 11030 ph: 516-562-4844 [office], 516-562-1135/4641 [lab] fax: 516-562-2866 **************************************************************************** *
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:29 EST