Description
Principle of the Assay: This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for CXCL1/KC has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CXCL1/KC present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for CXCL1/KC is added to the wells and binds to the combination of capture antibody-CXCL1/KC in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps, a substrate is added. A colored product TMB is formed in proportion to the amount of CXCL1/KC present in the sample. The reaction is terminated by addition of acid and absorbance is measured. A standard curve is prepared from seven CXCL1/KC standard dilutions and CXCL1/KC sample concentration determined.:Background/Introduction: CXCL1/GRO alpha, also known as KC in mouse and CINC-1 in rat, is a member of the CXC family of chemokines. Human CXCL1/GRO alpha is 107 amino acids (aa) in length with a predicted molecular weight of 11 kDa. The mouse and rat orthologs share 57% and 59% amino acid sequence identity with the human protein, respectively. Genome-wide analysis suggests that the protein encoded by the mouse Cxcl3 gene is homologous to human CXCL1/GRO alpha protein. Human CXCL1/GRO alpha can be cleaved into three isoforms CXCL1/GRO alpha (aa 4-73), CXCL1/GRO alpha (aa 5-73), and CXCL1/GRO alpha (aa 6-73) which show higher activity than the full length protein. CXCL1/GRO alpha is secreted by mast cells and macrophages whereupon it acts as a chemoattractant for neutrophils