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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.
Most mammal IgG class antibodies, including those from human are composed of two heavy and two light chains. Such light chains can be of kappa or lambda types, with an expected ratio of 3 kappas to one lambda. Both appear to play the same function in antibodies. Free light chains are usually rare in blood. However, several types of multiple myelomas will secrete free light chains, which can then accumulate in organs such as the kidney and cause severe damage, with lambda chains being the poorest diagnostic.
Knowledge of the type of light chain generated is also important for the creation and production of therapeutic antibodies. Techniques used for purification, such as protein L or anti species antibodies, can have different affinities for the two chains. The kappa light chain AlphaLISA detection kit allows for the detection of kappa light chain in human sera, plasma, and cell culture supernatants.
|Assay Target||Kappa light chain|
|Assay Target Class||Antibody|
|Experimental Type||In vitro|
|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.
This manual explains how to run the AlphaLISA no-wash human kappa light chain detection assay.