DEVELOPMENT AND SOME APPLICATIONS OF ENZYME-LINKED-IMMUNOSORBENT-ASSAY SYSTEM FOR MURINE MACROPHAGE INFLAMMATORY PROTEIN-2 (MIP-2)

WE attempted to establish an enzyme-linked Immunosorbent assay (ELISA) system by preparation of recombinant murine MIP-2 and its rabbit anti bodies, A fusion construct of MIP-2 to protein A was used to enable ea sy purification as well as the generation of a sufficiently large anti body response; The specificity of antibody was confirmed by Western bl otting analysis of 20-h conditioned medium from lipopolysaccharide (LP S)-stimulated RAW264.7 cells, a murine macrophage cell Line; antibody gave: a single band with a molecular weight of approximately 6000, whi ch is identical to that of murine MIP-2 reported previously, Biotin-st reptavidin sandwich ELISA could detect quantitatively MIP-2 at concent ration range of 20 to 1000 pg/ml, In some applications of this ELISA s ystem, time-related production of MIP-2 and inhibitory effect of dexam ethasone on its production have been demonstrated in LPS-stimulated RA W264.7 cells, Thus, ELISA system established in this study is consider ed to be a useful tool to study MIP-2 response in various inflammation models in mice.