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AlphaLISA technology allows the detection of molecules of interest in buffer, cell culture media, serum and plasma in a highly sensitive, quantitative, reproducible and user-friendly mode. In an AlphaLISA assay, a Biotinylated Anti-Analyte Antibody binds to the Streptavidin-coated Alpha 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 provokes 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.
The ubiquitin binding protein p62, or Sequestosome 1 (SQSTM1), is expressed in most cell types. Synonyms are p62 lck ligand, zeta interacting protein (ZIP, rat), A170 (mouse), OSIL, or PDB3. The human protein is 440 amino acids long. It contains an N-terminal protein binding domain (PB1) that interacts with different protein kinases and allows homopolymerization of p62. The domain is followed by several other protein-protein interaction domains and nuclear localization and export signals. Notably, the C-terminal ubiquitin association domain recognizes both mono and polyubiquitin and the LIR motif allows direct binding to the autophagosome-associated protein LC3. p62 plays an important role in the formation of intracellular protein aggregates, autophagy, and NF-κB signaling. Mutation of p62 is associated with Paget’s disease (abnormal bone turnover).
|Assay Target Class||Protein|
|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.
Protocol for AlphaLISA human p62 Detection Kit