An ELISA assay for murine interleukin-1β

• Published in: Journal of immunological methods Vol. 161; no. 2; pp. 257 - 264
• Main Authors: Newton, Robert C., Dowling, Randine, Daulerio, Andrea J., Culp, Steven
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 26.05.1993; Elsevier
• Subjects: Animals; Biological and medical sciences; ELISA; Enzyme-Linked Immunosorbent Assay; Fundamental and applied biological sciences. Psychology; Fundamental immunology; IL-1β; Immunoassay; Interleukin-1 - analysis; Male; Mice; Mice, Inbred C57BL; Molecular immunology; Rabbits; Sensitivity and Specificity; Techniques; PBS; Immunoassay; BSA; IL-1; GM-CSF; mIL-1; IgG; IL-1β; ELISA; Antibody; Cytokine; Rodentia; Rabbit; Interleukin 1β; Method; Lagomorpha; Vertebrata; Specificity; Mammalia; Mouse; Detection; ELISA assay; Comparative study
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(93)90301-M

An ELISA assay was developed for murine IL-1β (mIL-1β) using a polyclonal antibody generated in rabbits. The antibody was purified by affinity chromatography on protein A coupled to Sepharose followed by chromatography on mIL-1β coupled to Sepharose. The protein A and affinity purified populations were compared using radiolabeled mIL-1β and the results used to develop the conditions for the ELISA. The assay developed is sensitive to pg/ml concentrations of mIL-1β, is comparable in sensitivity to one which uses a hamster monoclonal antibody as the capture antibody, and can be used to detect IL-1β in peritoneal 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 without affecting assay sensitivity. This assay should prove useful for defining the presence and potential role for IL-1β in animal models of disease.