ELISA calls for help

Liuchuxin via methods%40net.bio.net (by liuchuxin from yahoo.cn)
Tue Apr 12 20:57:30 EST 2011


Dear sir/madam,
Sorry to interrupt 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 chemiluminescence ELISA, or the color ELISA?
                        
I am eager to your reply.
 
Best regards!
 
Chuxin Liu
Huazhong Agricultural University
Wuhan, China


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