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The Alpha SureFire® Ultra™ Multiplex kits allow the rapid, sensitive, and quantitative detection of two phosphoprotein targets in each well of an assay plate. This kit can be used in combination with any AlphaLISA SureFire Ultra assay to detect phospho-STAT6 (Tyr641) in a multiplexed assay.
Alpha SureFire assays are cellular assays used to detect and quantitate phosphoproteins for cell signaling, pathway analysis, and cellular kinase studies using homogeneous (no wash steps; no separation steps) Alpha technology. Terbium SureFire assays enable multiplexing for detection of two phosphoproteins in a single well, when used in conjunction with any AlphaLISA SureFire Ultra kit. This multiplex measurement is achieved by the use of two types of Alpha Acceptor beads that emit at distinct wavelengths (Terbium: 545 nm, Europium: 615 nm). The two distinct Alpha Acceptor beads report their binding to distinct antigens through their association with specific assay antibodies.
The Terbium “Alpha 545 Acceptor Beads” in the kit are conjugated with a “CaptSure2” agent, which binds a specific CaptSure2 tag on one of the assay antibodies. In combination with the biotinylated antibody provided, the 545 nm signal generated will report levels of the phosphoprotein of interest (Target 2). This kit is to be used in conjunction with the reagents in an AlphaLISA SureFire Ultra kit, which will report that target-of-interest by Europium emission at 615 nm (Target 1).
*These assays utilize the bead-based Alpha Technology, and requires an Alpha Technology-compatible plate reader capable of reading dual emission wavelengths. Refer to the “AlphaPlex Quick Start Guide” and the “AlphaPlex Assay Development Guide” to find guidance about filters and mirrors selection, instrument protocol and channels crosstalk correction.
Protein Analysis & Detection
|Assay Target Class
AlphaLISA SureFire Ultra Multiplex