The AlphaScreen® SureFire® total IκBα assay is a sandwich immunoassay for quantitative detection of IκBα (both phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be used as a normalization for phosphorylation studies. Please note that an AlphaScreen Protein A kit, which contains the Alpha Beads necessary to run this assay, needs to be purchased separately in addition to the AlphaScreen SureFire kit.
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Alpha SureFire® kits can be used for:
In the AlphaScreen® SureFire® assay, Donor beads are coated with streptavidin to capture one of the antibodies, which is biotinylated. Acceptor beads are coated with Protein A to immobilize the other antibody. In the presence of target protein, the two antibodies bring the Donor and Acceptor beads close together, generating signal. The amount of light emission is directly proportional to the amount of protein present in the sample.
|Assay Target Class||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaScreen SureFire|
|Quantity in a Package Amount||10000.0 Units|
|Shipping Condition||Blue Ice|
|Unit Size||10,000 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.
Screening kinase activities in cell-based assays offers advantages over the more traditional biochemical approach of using a purified recombinant enzyme to phosphorylate a substrate, since a cell-based assay gives information on a compound’s activity in a more biological context. However, getting initial results with the AlphaScreen® SureFire® assay is highly dependent on optimal cell culture conditions, and often requires that multiple parametersbe optimized in the first set of experiments.
Cell lines validated for use with AlphaScreen® SureFire® kits