ELISA assays vs immunofluor

Danny Altschuler dla at galactose.mc.duke.edu
Thu May 22 14:17:39 EST 1997




General question: I have a mix population of stable transfected cells. By
immunofluorescence positive expression represents about 1% of the
population. To get independent clones I will need to screen at least 100
independent clones. I was planning to use the same antibody (btw is
HA11monoclonal) to do ELISA type of assays in 96well plates using a
peroxidase colorimetric assay and read on a plate reader.
Is the system sensitive enough?\
Can I assume that the protocol I used for Immunofluorescence will work for
ELISA? (4% PF, 0.5% Triton etc)
Does anyone have experience in screening large number of clones? I should
point out that I have 6 mutants, so the number of clones to screen will be
at least 600.
Can I have the 96well plates in medium with 10% DMSO and freeze them down
to -70, during screening time?
Any suggestions, please e-mail to
altschul at med.unc.edu
Thanks
Danny 



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