I am developing an ELISA assay (96 wells plates) for detection of mouse resistin from 3T3-L1 cells. I have diluted my kit according to the instructions from the developer (R&D Systems) but are experiencing problems with the standards when I run an experiment. The antibodies seem to work properly because there is a clear presence of resistin in my sample. However, the standards are pretty much undetectable, even the high one of 1000 pg/ml. The standards are diluted according to the instructions (and my calculations are double checked by my lab partner) so I am wondering what could be the problem with my standard? So far I am suspecting that it may be something wrong with the standards that I have gotten from the manufacturer. Has anyone experienced anything similar?
I am thankful for any advice/answer!