The AlphaLISA® Human Matrix Metalloproteinase-9 (MMP9) Detection Kit is designed for detection and quantitation of human MMP9 in serum, buffered solution or cell culture medium in a homogeneous (no-wash steps, no separation steps) assay. The kit was designed to detect both MMP9 and pro-MMP9.
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Matrix Metalloproteinase-9 (MMP9) or gelatinase B is a 64-82 kDa, zinc- and calcium-dependent, endopeptidase secreted by normal alveolar macrophages and granulocytes. MMP9 is secreted as a pro-enzyme (pro-MMP9) and is activated by proteolytic cleavage by serine proteases including other MMPs. MMP9 is believed to act synergistically with MMP1 interstitial collagenase in the degradation of fibrillar collagens. MMP9 might contribute to the pathogenesis of joint destruction. MMP9 could also play a role in bone osteoclastic resorption and cancer metastasis. MMP9 constitutes a useful biomarker of disease status in arthritis and certain cancers.
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
|Assay Target Class||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 assay points|
Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.
The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.
Enzyme-linked Immunosorbent Assay is the most widely Kits adopted method for detection and quantification of cytokines and other biomarkers. This traditional technology offers good,selectivity, sensitivity and assay versatility; however, it has certain disadvantages such as limited dynamic range and low throughput due to the numerous wash steps.