描述：
描述
The Human Matrix Metalloproteinase-9 (MMP-9, MMP9) ELISA research-use-only kit is to be used for the quantitative determination of MMP-9 in samples (see sample types indicated) using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant forms of this target.
Performance characteristics
• Sensitivity: <10 pg/mL
• Standard curve range: 23.5‒1,500 pg/mL
• Sample type(s): serum, plasma, cell culture supernatant
• Specificity: natural and recombinant human MMP-9
• Cross-reactivity: see kit manual for cross-species and/or cross-target reactivity
• Sample volume: 100 μL
• Total assay time: 4 hours
Rigorous validation
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Principle of the method
The human MMP-9 solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, binding to the target on a different epitope from the capture antibody. A conjugated enzyme has been incorporated into the assay. After incubation and washing steps to rid the microplate of unbound substances, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.
Target information
Matrix metalloproteinase (MMP) is a class of zinc-dependent proteolytic enzymes that are responsible for the degradation of extracellular matrix (ECM) proteins. To date, more than 20 different human MMPs have been cloned and characterized. According to their structure, substrate specificity, and cellular localization, they can be classified into several subgroups, gelatinases (MMP-2 and MMP-9), interstitial collagenases (MMP-1, MMP-8, MMP-13, and MMP-18), stromelysins (MMP-3 and MMP-10), matrilysin (MMP-7), metalloelastases (MMP-12), and membrane type MMPs (MT-MMPs). The expression of MMPs is transcriptionally regulated by growth factors, hormones, cytokines, and cellular transformation. Most MMPs, except MT-MMPs, are synthesized as pre-proenzymes and secreted as inactive zymogens into the extracellular matrix, where subsequent activation results in cleavage of the proenzymes into the active species. The proteolytic activities of MMPs are precisely controlled during activation from their precursors and inhibition by endogenous inhibitors, α-macroglobulins and tissue inhibitors of metalloproteinases (TIMPs).
MMP-9, also known as gelatinase B, 92 kDa gelatinase, and 92 kDa type IV collagenase, represents the largest and most complex member of MMP family. MMP-9 comprises five distinct domains: a propeptide domain which is cleaved upon activation, a gelatin-binding domain consisting of three contiguous fibronectin type II units, a catalytic domain containing the zinc-binding motif, a proline-rich linker region, and a carboxyl terminal hemopexin–like domain. Like other MMPs, MMP-9 is first synthesized as an inactive proenzyme. Activation of proMMP-9 is mediated by plasminogen activator/plasmin system and other MMPs. The regulation of MMP-9 activity is also controlled through TIMPs.
MMP-9 is produced by many types of cells, such as monocytes, macrophages, neutrophils, keratinocytes, fibroblasts, osteoclasts, chondrocytes, endothelial cells, and various tumor cells. Expression of MMP-9 is regulated by several cytokines and growth factors, including interleukins, interferons, EGF, NGF, FGF, VEGF, PDGF, TNF-α, TGF-β, the extracellular matrix metalloproteinase inducer EMMPRIN and also osteopontin. Upon activation, MMP-9 acts on a broad range of substrates including gelatin, collagen types IV, V, XI, and XVI, elastin, decorin, fibrillin, laminin and also activates growth factors like proTGF-β and proTNF-α. Physiologically, MMP-9 in coordination with other MMPs, play a role in normal tissue remodeling events such as neurite gowth, embryonic development, angiogenesis, ovulation, mammary gland involution and wound healing. MMP-9 with other MMPs is also involved in osteoblastic bone formation and/or inhibits osteoclastic bone resorption.
Related links
Learn more about ELISA kits
Learn more about other immunoassays
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