Matrix metalloproteinase-14 is an enzyme that in humans is encoded by the MMP14 gene. It is mapped to chromosome 14q11-q12. Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive pro-proteins which are activated when cleaved by extracellular proteinases. However, the protein encoded by this gene is a member of the membrane-type MMP (MT-MMP) subfamily; each member of this subfamily contains a potential transmembrane domain suggesting that these proteins are expressed at the cell surface rather than secreted.
Sample Type
Cell culture supernates, serum, plasma (heparin or EDTA).
Intended Use
Sandwich High Sensitivity BioAssay™ ELISA kit for Quantitative Detection of Human MMP-14.
Range
156pg/ml-10000pg/ml
Specificity
Recognizes natural and recombinant Human MMP-14. There is no detectable cross-reactivity with other relevant proteins.
Test Principle
This Human MMP-14 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for MMP-14 has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli; Immunogen sequence: A21-S538) and test samples are added to the wells, a biotinylated detection specific for MMP-14 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 Human MMP-14 amount of sample captured in plate.
Kit Components
96-well plate precoated with anti-Human MMP14 antibody, 1x plate Lyophilized recombinant Human MMP14 standard, 2x10ng/tube Biotinylated anti-Human MMP14 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 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.