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. The standard product used in this kit is natural, isolating from human MMP-8. The detected MMP-8 includes zymogen and active enzyme.
Sample Type
Cell culture supernates, serum and plasma (heparin)
Intended Use
For quantitative detection of rat MMP-8 in cell culture supernates, serum and plasma(heparin).
Specificity
Natural and recombinant rat MMP-8
Test Principle
This rat MMP-8 BioAssay™ ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for MMP-8 has been precoated onto 96-well plates. Standards(NSO, L21-P466) and test samples are added to the wells, a biotinylated detection 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 is added and unbound conjugates are washed away with PBS or TBS buffer. HRP substrate TMB are used to visualize HRP enzymatic reaction. TMB is 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 rat MMP-8 amount of sample captured in plate.
Kit Components
96-well plate precoated with rat MMP-8 antibody, 1 Lyophilized recombinant rat MMP-8 standard, 10ng/tube×2 Biotinylated rat MMP-8 antibody, 130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100) Sample diluent buffer, 30ml Antibody diluent buffer, 12ml ABC diluent buffer, 12ml TMB color developing agent, 10ml TMB stop solution, 10ml
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
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