Higher sensitivity
James W. Jacobberger (jwj@po.CWRU.Edu)
Wed, 2 Aug 1995 12:27:07 -0400
+
Steve
If you look for Srivastava et al, Cytometry 13:711-721, 1992,
you'll see streptavidin staining described for low expression
intracellular molecules. This was for intracellular molecules
but the principles should be the same. For the surface, you
may be able to do a little better since your background may
be less. The current estimate for the lowest expression cell
line used in that paper is ~1000-2000 molecules.
Another approach may be to stain your cells while live, then
fix with low level formaldehyde, then treat with detergent to
release much of the cyotplasmic components (autofluorescence
goes down considerably). See Clevenger 1986, Cytometry for
this type of procedure. Another approach is to use more than
one epitope for primary staining. In that way, we could
increase the specific fluorescence ~1.9 times in some instances
(again, intracellular antigens).
The other obvious choice is to use phyocobili protein conjugates.
The reference for Clevenger is Cytometry 6:208-214, 1985.
--
James W. Jacobberger
Case Western Reserve Univ.
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