Matrix metalloproteinase 8 (MMP8) also called neutrophil collagenase. Neutrophil collagenase, a member of the family of matrix metalloproteinases, is distinct from the collagenase of skin fibroblasts and synovial cells in substrate specificity and immunologic crossreactivity. MMP8, an enzyme that degrades fibrillar collagens imparting strength to the fetal membranes, is expressed by leukocytes and chorionic cytotrophoblast cells. The human neutrophil collagenase (HNC) cDNA clone has been sequenced and shown to encode a 467-residue protein. Neutrophil collagenase has been found to possess 57% identity with the deduced protein sequence for fibroblast collagenase with 72% chemical similarity. Certain regions of the molecule, including the putative zinc-binding region, are highly conserved. When compared with the published sequence for fibroblast collagenase, neutrophil collagenase contains four additional sites for glycosylation. Sample Type: Cell culture supernates, serum, plasma (heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Mouse MMP-8. Sensitivity: <10pg/ml Range: 156pg/ml-10000pg/ml Specificity: Recognizes natural and recombinant Mouse MMP-8. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Mouse MMP-8 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A a monoclonal antibody from rat specific for MMP-8 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: F21-S465) and test samples are added to the wells, a biotinylated detection a biotinylated polyclonal antibody from goat specific for MMP-8 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Mouse MMP-8 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Mouse Mmp8 antibody, 1x plate Lyophilized recombinant Mouse Mmp8 standard, 2x10ng/tube Biotinylated anti-Mouse Mmp8 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
