Alpha SureFire® Ultra™ Multiplex p-Akt 1/2/3 (S473) (Eu) + Total Akt1 (Tb) Kit, 10,000 Assay Points

The Alpha SureFire® Ultra™ HV Multiplex p-Akt 1/2/3 (Ser473) + Total Akt 1 assay kit is used to measure both the phosphorylation (Ser473) and total levels of endogenous Akt in cellular lysates. The kit measures phosphorylation on Ser473 of Akt 1/2/3, and total levels of Akt 1 only. 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 (Ser473) activation by either recombinant or endogenous receptors, and can be applied to primary cells.

The 615nm (Eu) signal corresponds to the phosphorylated Akt 1/2/3 (Ser473) analysis, and the 545nm (Tb) signal corresponds to the total Akt 1 analysis.

This kit has been formulated to provide improved signal:background (i.e. S:B) assay windows, and to perform without interference in the presence of extraneous antibodies.

• No-wash steps, no separation steps
• Multiplexing assay
• Sensitive detection
• No interference from exogenous antibodies
• Results in less than 3 hours


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In this assay, the Alpha 615 Acceptor bead is directly-conjugated with an antibody to the phosphorylated site on the target protein. The Alpha 545 Acceptor bead is coated with the CaptSure agent, which binds the CaptSure tagged anti-total target protein antibody. The Alpha Donor bead binds the biotinylated anti-total target protein antibody.

The antibodies used in this kit are identical to those used in the individual phospho- (Ser473) and total- Akt AlphaLISA SureFire Ultra kits (ALSU-PAKT-B and ALSU-TAKT1-A catalog numbers), although their orientation of bead attachment may vary between kits.