[Immunology] ELISA calls for help

Liuchuxin via immuno%40net.bio.net (by liuchuxin from yahoo.cn)
Thu Apr 14 07:42:53 EST 2011


Dear sir/madam,
Sorry to disturb you! I am a postgraduate in China, and I get in trouble in
ELISA.  In ELISA procedure, I am confused at the final step-data processing. In
my work, I used luminol as substrate to generate chemiluminescentsignals, rather
than TMB to bring color changes. However, in my indirect ELISA to detect
antibodies in mouse serum, the negative sample or even the blank sample will
always produce quiet strong signals, eg. 22366, 30512, 31290 for the
chemiluminescence density values of three antiserum samples, 22621 and 22152 of
the two negative samples, 22435 of the blank one (no coating reagent or
substrate added). The difference between the antiserum and control serum or
blank sample is tiny, how can I diminish the background signal and how to assess
the antibody titers in this condition? What are the advantages and disadvantages
of the fluorescent ELISA, or the color ELISA?
                       
I am eager to your reply.
 
Best regards!
 
Chuxin Liu
Huazhong Agricultural University
Wuhan, China


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