Is this always a better idea? I did an experiment a while back in
which I used indirect ("sandwich") staining with an antibody against MHC
class I. By the time I got around to the analysis, the light scatter prop-
erties of the cells that were positive for my antibody were all out of whack,
as if I had stimulated them. It reminded me of a technique I once used for
stimulating T cells, in which you added anti-CD3-biotin and then streptavidin.
So, if cross-linking the antigen in question would stimulate your
cells, perhaps you want to use direct-labeled antibodies. Comments?
- John Ladasky
ladasky@leland.stanford.edu