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One antibody is specific to the ß-secretase cleavage site at the N-terminus: mouse monoclonal antibody, clone number 82E1. The second antibody is specific to the C-terminus: mouse monoclonal antibody, clone number 12F4.
Amyloid beta (Aß) is a short peptide derived from the proteolysis of a larger transmembrane molecule, the amyloid precursor protein (APP). The ß- and γ-secretases cleave the respective N- and C-terminal ends of the Aß sequence, liberating the Aß peptide from APP. Aß40 is the major species of Aß produced by neurons and other cells, and accounts for over 70% of total Aß produced, while the remaining 10-20% is comprised of the longer Aß42, and other species. Aß42 has a greater propensity to form aggregates or fibrils and also has a greater neuronal toxicity in tissue culture models than Aß40, implying that Aß42 is a more important factor in Alzheimer's disease (AD) pathogenesis and plaque formation. Levels of Aß42 in cerebrospinal fluid are decreased in the majority of AD subjects (probably due to its aggregation into plaques), making it an important biomarker for this 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 Class||Peptide|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Therapeutic Area||Central Nervous System|
|Unit Size||500 assay points|
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.