HSA Eliza (fwd)

Hao Xiao 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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