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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.
Collagen IV, as one of many extracellular matrix proteins, is produced by fibroblasts, endothelial cells, and epithelial cells. It is a heterotrimeric molecule containing two α1-like and one α2-like chain. The chain compositions of collagen IV are associated with six genes (COL4A1, COL4A2, COL4A3, COL4A4, COL4A5, COL4A6) suggesting that the exisistence of additional alpha-chains to collagen structures and functions. Collagen Type IV is found mainly in the basement membranes of many tissiues and organs including liver, kindney, and skin. It functions to maintian the structural integrity of cells, tissues, and organs. Structural alterations of collagen IV is associated with many diseases such as nephritic syndrome, hemoptysis, liver fibrosis, subepidermal blistering diseases in the skin, and diabetic nephropathy. Concentrations of collagen IV in serum correlate with alcoholic liver disease.
|Assay Target||Collagen IV|
|Assay Target Class||Collagen|
|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 collagen IV detection assay.