An ELISA assay was developed for murine IL-1beta (mIL-1beta) using a p
olyclonal antibody generated in rabbits. The antibody was purified by
affinity chromatography on protein A coupled to Sepharose followed by
chromatography on mIL-1beta coupled to Sepharose. The protein A and af
finity purified populations were compared using radiolabeled mIL-1beta
and the results used to develop the conditions for the ELISA. The ass
ay developed is sensitive to pg/ml concentrations of mIL-1beta, is com
parable in sensitivity to one which uses a hamster monoclonal antibody
as the capture antibody, and can be used to detect IL-1beta in perito
neal washings or tissue lysates from either mouse or rat. There is no
cross reaction with any cytokine tested. The use of ELISA enhancement
kits can increase the resolution at the lower concentration ranges wit
hout affecting assay sensitivity. This assay should prove useful for d
efining the presence and potential role for IL-1beta in animal models
of disease.