HSA Eliza (fwd)
haoxiao at cc.UManitoba.CA
Wed Oct 26 14:22:04 EST 1994
Sorin Damian writes:
> From BIOSCI-REQUEST at net.bio.net Wed Oct 26 03:49:19 1994
> To: immunology at net.bio.net
> From: sorind at crash.cts.com (Sorin Damian)
> Subject: HSA Eliza
> Date: Wed, 26 Oct 1994 05:47:04 GMT
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> Problem: too much nonspecific binding during Eliza of Human Serum Albumin
> (HSA) in hepatocyte culture.
> I am using the HSA MoAfrom sigma , the secondary is a peroxidase
> conjugated IgG. I have far FAR to much nonspecific binding of the primary
> anti HSA MoA to the plastic (or maybe the conc ratio between
> antigen/primary , or btw primary and secondary is not the right one...
> Any help ?
> I can give more details of the experimental approach for those
> able/willing to post their experience.
> Any one out there working on HSA quantitation by ELIZA?
If so, it would be good for direct assay. Biotinylation of your standard HSA
and adsorbed you mAb to the plate and the sample HSA will compete with the
biotin-HSA followed by avidin-HRP probing. Biotinylation of HSA may partially
destroy the immunodeterminant of HSA specific for your mAb, but it still
provide rapid and sensitive detection, believe me. biotin-avidin-HRP
interaction is much faster than that for 1st Ab and 2nd Ab-HRP.
Ph.D student in Pharmacolgy and Toxicology
haoxiao at cc.umanitoba
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