The AlphaLISA® immunoassay kit for detection and quantitation of human Amyloid β 1-40 (Aβ 1-40) in cerebrospinal fluid (CSF), cell culture supernatants, and other sample types allows for fast, reproducible, and sensitive detection without the need for time-consuming wash steps or complicated assay development.
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One antibody is specific to amino acids 17-24 (epitope VFFAE): mouse monoclonal antibody, clone number 4G8. 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 Aβ 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||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.
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.