Recently I have been working with detecting serum IL12 by ELISA. Based on that result, the sera from healthy mice have around 10000~20000pg/mL IL12p40/p70. But when I searched the literature, IL-12p40 is around 200pg/mL. The coating antibody and detection antibody I used can both recognize p40 and p70. The standard curve looked fine. I diluted the serum by 30 fold, and I could still detect the OD signal. And the negative control (buffer alone) had a low background.
I am wondering whether it is because the antibody can detect both p40 and p70. Still, the concentration of p40 plus p70 is too high. And does anybody know what specific antibody should I use to only detect IL12?
Dear Daxing
Please review http://www.jimmunol.org/content/162/3/1582.full.pdf
Hakeem Sam and Mary M. Stevenson have described specific antibodies that may resolve your issues.
Thanks! This paper has similar basal serum IL12p40 level with my result(10-20ng/mL). And we used the same antibodies(C15.6/C17.8). But for most papers, when they measured IL12p40, the basal level is around 200pg/mL. Some people use C15.1/sheep polyclonal antibody for IL12p40. I don't know whether this can explain it.
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