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Mediators of Inflammation
Volume 5 (1996), Issue 3, Pages 206-209
http://dx.doi.org/10.1155/S0962935196000294

Development and some applications of enzyme-linked immunosorbent assay system for murine macrophage inflammatory protein-2 (MIP-2)

1Department of Human Science, Faculty of Medicine, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama 930-01, Japan
2Department of japanese Oriental Medicine, Faculty of Medicine, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama 930-01, Japan
3Department of Virology, Medical School, Nagoya City University, Mizuho-machi, Nagoya 467, Japan

Copyright © 1996 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

We attempted to establish an enzyme-linked immunosorbent assay (ELISA) system by preparation of recombinant murine MIP-2 and its rabbit antibodies. A fusion construct of MIP-2 to protein A was used to enable easy purification as well as the generation of a sufficiently large antibody response. The specificity of antibody was confirmed by Western blotting analysis of 20-h conditioned medium from lipopolysaccharide (LPS)-stimulated RAW264.7 cells, a murine macrophage cell line; antibody gave a single band with a molecular weight of approximately 6000, which is identical to that of murine MIP-2 reported previously. Biotin–streptavidin sandwich ELISA could detect quantitatively MIP-2 at concentration range of 20 to 1000 pg/ml. In some applications of this ELISA system, time-related production of MIP-2 and inhibitory effect of dexamethasone on its production have been demonstrated in LPS-stimulated RAW264.7 cells. Thus, ELISA system established in this study is considered to be a useful tool to study MIP-2 response in various inflammation models in mice.