The AlphaLISA® SureFire® Ultra™ Human and Mouse phospho-AKT 1/2/3 (Thr450)/Total AKT1 assay kit is used to measure both the phosphorylation of AKT 1/2/3 and total levels of endogenous AKT 1 in cellular lysates. The assay is an ideal system for the screening of modulators of receptor activation (e.g. agonists and antagonists) as well as agents acting intracellularly, such as small molecule inhibitors of signal transduction. The assay will measure AKT 1/2/3 phosphorylation by either recombinant or endogenous receptors and can be applied to primary cells.
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Protein kinase B (PKB) also known as AKT1 AKT2 and AKT3 encode the RAC alpha, beta, and gamma serine/threonine protein kinases respectively. AKT1 is involved in cellular survival pathways, by inhibiting apoptotic process. It is also able to induce protein synthesis pathways and is therefore a key signaling protein in the cellular pathways that lead to skeletal muscle hypertrophy and general tissue growth. Akt2 is an important signaling molecule in the insulin signaling pathway. AKT3 appears to be predominantly expressed in the brain.
The AlphaLISA SureFire Ultra Multiplex Human and Mouse Phospho-AKT 1/2/3 (Thr450)/Total AKT1 Detection Kit is used to measure phosphorylated AKT 1/2/3 and total levels of AKT1 in cellular lysates in a multiplexing Alpha no-wash assay. No specially engineered cell lines are required. The 615 nm (Eu) signal corresponds to the phosphorylated AKT 1/2/3, and the 545 nm (Tb) signal corresponds to the total AKT1 analysis. This kit has been formulated to provide superior signal:background assay windows and to perform without interference in the presence of extraneous antibodies, making it amenable to the study of therapeutic and blocking antibodies.
In this assay, the Alpha 615 Acceptor bead is coated with the CaptSure™ antibody, which binds the CaptSure-tagged anti-phospho target antibody. The Alpha 545 Acceptor bead is coated with the CaptSure2™ agent, which binds the CaptSure2 tagged anti-total target protein antibody. The Alpha Donor bead binds the biotinylated anti-total target protein antibody.
|Assay Points (200-50,000)||50000 Assay Points|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA SureFire Ultra Multiplex|
|Shipping Condition||Blue Ice|
|Target Species||Human, Mouse|
|Therapeutic Area||Cardiovascular, Oncology|
|Unit Size||50,000 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.
Product brochure for the Alpha Technology, a versatile, no wash, homogeneous assay technology that's suitable for a broad range of applications.
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