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Frataxin is expressed as a 210 AA, 23 kDA protein from the FXN gene located on chromosome 9. Upon expression, the FXN protein is directed to the mitochondrion by its 41 AA N-terminal mitochondrial targeting sequence. In the mitochondrion, the protein is cleaved by the mitochondrial processing peptidase (MPP) to its intermediate form of 42 – 210 AA. Later on, MPP cleaves the protein to its mature form of 81-210 AA. Although the function of the FXN protein is not clearly defined, it is thought to be vitally important for Iron-Sulfur cluster biogenesis, heme biosynthesis, and chelation and transportation of iron specifically involved with the mitochondria. The Frataxin protein is the primary culprit for a debilitating neurodegenerative disease called Friedreich’s Ataxia. Due to similarities in the diseases, there is some evidence to suggest that FXN may be involved in other neurodegenerative diseases such as Parkinson’s, Multiple Sclerosis, and Amyotrophic Lateral Sclerosis. This AlphaLISA kit has been designed for the detection and quantification of FXN from cell and tissue lysates.
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||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Quantity in a Package Amount||1.0 per pack|
|Shipping Condition||Blue Ice|
|Therapeutic Area||Central Nervous System|
|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.