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The kit was designed to equally detect free Troponin I, Troponin IC complex, or Troponin ITC complex. Troponin I is detected using two mouse monoclonal antibodies: clone numbers M18 and 267.
Troponin is one of the main proteins involved in muscle contraction and consists of three subunits: troponin C, involved in calcium binding; troponin I, the inhibitory subunit; and Troponin T, the tropomyosin binding subunit. Cardiac TnI (cTnI) is a well known serum biomarker of cardiac muscle injury, especially of myocardial infarction. Once released in the blood stream, cTnI is mainly in the binary complex of cTnI and cTnC (42 kDa), but also is present as the free form (24 kDa) and cTnI-cTnT-cTnC or ITC complex (78 kDa). Also, it has been found that healthy subjects having cTnI blood concentrations over the 99th percentile are at a higher risk of cardiovascular disease.
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||cardiac Troponin I|
|Assay Target Class||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 assay points|
The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.
Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.
For the detection of three biomarkers in complex sample matrices, the AlphaLISA and Electrochemiluminescent (ECL) assay technologies were shown to have similar: Assay windows (linear dynamic range), Lower and upper detection limit, Intra-and inter-assay precision (lower % CV) The advantages of using AlphaLISA over ECL are: Shorter total assay duration No wash steps No shaking Lower sample volume requirement for equivalent performance Less expensive instrument and plates required
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.