The AlphaLISA® SureFire® Ultra™ Total STAT5 assay is a sandwich immunoassay for quantitative detection of STAT5 (both phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be used as a normalization for phosphorylation studies.

The HV (high volume) kit contains reagents to run 100 wells in 96-well format, using a 60 µL reaction volume. The 500 point kit contains reagents to run 500 wells in 384-well format, using a 20 µL reaction volume. The 10,000 point kit contains reagents to run 10,000 wells in 384-well format, using a 20 µL reaction volume. The 50,000 point kit contains reagents to run 500 wells in 384-well format, using a 20 µL reaction volume.

In the AlphaLISA® SureFire® Ultra™ assay, Donor beads are coated with streptavidin to capture one of the antibodies, which is biotinylated. Acceptor beads are coated with a proprietary CaptSure™ agent that immobilizes the other antibody, labeled with a CaptSure™ tag. 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.

AlphaLISA® SureFire® Ultra™ kits are compatible with:

• Cell and tissue lysates
• Antibody modulators
• Biotherapeutic antibodies

Alpha SureFire® kits can be used for:

• Cellular kinase assays
• Receptor activation studies
• Screening