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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.
Heme oxygenases are enzymes that are involved in the degradation of heme. They cleave the heme ring to create billiverdin, which then becomes bilirubin due to the action of bilirubin reductase. This cascade allows for the removal of heme in damaged red blood cells as well as heme bound irreversibly to molecules such as carbon monoxide. Heme oxygenases have been characterized as markers for oxidative stress, metabolic disorders and inflammation. Heme oxygenase 2 is present mainly in the brain. Its role appears to be protective against oxidative stress. Production of carbon monoxide from the cleavage of the heme ring seems to play a protective role against malaria by poisoning the hemoglobin essential for the parasite growth.
|Assay Target Class||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||500 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.
This manual explains how to run the AlphaLISA no-wash human heme oxygenase 2 detection assay.