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EK000002-EK0464: Human MMP-8 ELISA Kit
Range 156pg/ml-10,000pg/ml
Sensitivity < 10 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human MMP-8 in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
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.1 The human neutrophil collagenase (HNC) cDNA clone has been sequenced and shown to encode a 467-residue protein.2 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.3 The standard product used in this kit is natural, isolating from human MMP-8. The detected MMP-8 includes zymogen and active enzyme.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Wang, H.; Parry, S.; Macones, G.; Sammel, M. D.; Ferrand, P. E.; Kuivaniemi, H.; Tromp, G.; Halder, I.; Shriver, M. D.; Romero, R.; Strauss, J. F., III Functionally significant SNP MMP8 promoter haplotypes and preterm premature rupture of membranes (PPROM). Hum. Molec. Genet. 13: 2659-2669, 2004.
2. Devarajan, P.; Mookhtiar, K.; Van Wart, H.; Berliner, N. Structure and expression of the cDNA encoding human neutrophil collagenase. Blood 77: 2731-2738, 1991.
3. Hasty, K. A.; Pourmotabbed, T. F.; Goldberg, G. I.; Thompson, J. P.; Spinella, D. G.; Stevens, R. M.; Mainardi, C. L. Human neutrophil collagenase: a distinct gene product with homology to other matrix metalloproteinases. J. Biol. Chem. 265: 11421-11424, 1990.
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