The AlphaLISA® Human Tissue Plasminogen Activator (tPA) Detection Kit is designed for detection and quantitation of human tPA 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 tPA and pro-tPA.
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Tissue Plasminogen Activator (tPA) is a 59 kDa secreted serine protease that converts the proenzyme plasminogen to plasmin, a fibrinolytic enzyme. It is synthesized in numerous tissues and is the principal endogenous activator of plasminogen in blood. It plays a role in cell migration and tissue remodeling. Increased enzymatic activity causes hyperfibrinolysis, which manifests as excessive bleeding; decreased activity leads to hypofibrinolysis that can result in thrombosis or embolism. Rapid fluctuations in tPA concentration can be observed in response to exercise, venous occlusion, alcohol, and drugs, such as anabolic steroids. Individuals who do not show increased tPA activity when exposed to some of these stimuli, may be at risk for deep vein thrombosis.
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.