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).

Formats:

• Our 100 assay point kit allows you to run 100 wells in 96-well format, using a 60 µL reaction volume (30 µL of lysate).
• Our 500 assay point kit allows you to run 500 wells in 384-well format, using a 20 µL reaction volume (10 µL of lysate).
• Our 10,000 assay point kit allows you to run 10,000 wells in in 384-well format, using a 20 µL reaction volume (10 µL of lysate).
• Our 50,000 assay point kit allows you to run 50,000 wells in in 384-well format, using a 20 µL reaction volume (10 µL of lysate).

Features:

• No-wash steps, no separation steps
• ELISA alternative technology
• Highly sensitive detection
• Results in less than 3 hours
• Half the time of an ELISA assay

*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.