Since this is my first post in here, HI ALL
As a med student doing a year of research I might need a bit of help
I'm setting up an ELISA for measuring human IL-35 (EBI3/p35 heterodimer) however until now I haven't been able to detect a signal when I read my plates. My initial setup has been coating the plates with a anti-human-p35 mAb (mouse, R&D Systems MAB1570) and detecting with a biotinylated anti-human-IL27 (antibody against both EBI3 and p28) (goat, R&D Systems #BAF2526). As a standard I have used a human IL-35:Fc Chimera protein (Enzo Life Sciences #ALX-522-140-C010).
I haven't run any samples yet just testing my ability to detect my standard (1000pg/ml) which until know haven't worked.
I'm pretty confident about my capturing antibody however my detection I'm not sure about since I would have prefered using a biotinylated mAb against EBI3. Does anyone have any suggestions how to solve this problem og maybe even have a matched pair already working?
Thx in advance
Institute of Biomedicine