I use 1% BSA for blocking ELISA plates. However, even after blocking for 3 hours, I still get background noise for some of the cytokines but not all. What can be the reason for this background noise and how can I tackle this problem.
Probably because this is obviously serum derived and contains low levels of cross-reacting antigens whether these be bovine cytokines or other antigens. You could try the Pierce protein-free blocking agent which has overcome this problem in tricky assays that we have developed, available through Thermo Fisher.