dual staining
Gina Pighetti (gmp109@email.psu.edu)
Thu, 19 Sep 1996 15:03:31 -0400
Hi, my name is Gina Pighetti and I am a Ph.D. student at Penn State. I am
interested in dual staining for lymhocyte surface markers in bovine dairy
cattle. Unfortunately, the monoclonal antibodies are not conjugated to
fluorescent markers so we must do indirect dual staining for both markers.
I was wondering what the best method would be. Should we stain for a single
monoclonal antibody (CD2) and its secondary (FITC) and then the next
monoclonal antibody (IL2Ra). Or should we add both monoclonal antibodies at
the same time, incubate, wash, and then add the secondary antibodies at the
same time? Is there a problem with high backgrounds with a double indirect
stain? Do we need a blocking step? We normally use phosphate buffered
saline with 2% fetal bovine serum for flow cytometery, is this a problem?
Any suggestions or comments on this would be greatly appreciated!! Thank
you, Gina
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
