Revvity, ASBF-PST6-B10K, AlphaLISA Biotin-Free High Performance Human Phospho-STAT6 (Tyr641) Detection Kit, 10,000 assay points

Overview
Signal Transducer and Activator of Transcription 6 (STAT6) is a key component of the JAK/STAT pathway, primarily activated by interleukin-4 (IL-4) and interleukin-13 (IL-13). Upon activation, STAT6 dimerizes and translocates to the nucleus, regulating genes involved in T helper 2 (Th2) immune responses, allergic inflammation, and cell survival. Dysregulation of STAT6 is implicated in asthma, autoimmune diseases, and cancers such as lymphomas and colorectal cancer, where it promotes tumor progression.
The AlphaLISA SureFire Biotin-Free High Performance Human Phospho-STAT6 (Tyr641) Detection Kit is a sandwich immunoassay for the quantitative detection of phospho-STAT6 (Tyr641) in cellular lysates, using Alpha Technology.
Formats:

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 enough reagents to run 500 wells in 384-well format, using a 20 μL reaction volume.
The 10,000-point kit contains enough reagents to run 10,000 wells in 384-well format, using a 20 μL reaction volume.
The 50,000-point kit contains enough reagents to run 50,000 wells in 384-well format, using a 20 μL reaction volume.

AlphaLISA SureFire Ultra kits are compatible with:

Cell and tissue lysates
Antibody modulators
Biotherapeutic antibodies

AlphaLISA SureFire Ultra kits can be used for:

Cellular kinase assays
Receptor activation studies
High-throughput screening for preclinical studies
How it works
Phospho-AlphaLISA SureFire Ultra Biotin Free assay principle
The Phospho-AlphaLISA SureFire Ultra Biotin Free assay measures a protein target when phosphorylated at a specific residue.
The assay uses two antibodies which recognize the phospho epitope and a distal epitope on the targeted protein. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure™ agent to specifically immobilize the assay specific antibody, labeled with a CaptSure tag. Donor beads are coated with a proprietary CaptSure 3 agent to capture one of the detection antibodies, which is labeled with CaptSure 3 tag. In the presence of phosphorylated protein, the two antibodies bring the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, allowing the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of phosphoprotein present in the sample.

 

Phospho-AlphaLISA SureFire Ultra Biotin Free two-plate assay protocol
The two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well Optiplate™ plate before the addition of Phospho-AlphaLISA SureFire Ultra Biotin Free detection reagents. This protocol permits the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.

Phospho-AlphaLISA SureFire Ultra Biotin Free one-plate assay protocol
Detection of Phosphorylated target protein with AlphaLISA SureFire Ultra Biotin Free reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol allows for miniaturization while maintaining robust AlphaLISA SureFire Ultra quality.
Application-Cell Signaling
Automation Compatible-Yes
Brand-AlphaLISA SureFire Biotin-Free
Detection Modality-Alpha
Product Group-Kit
Protocol Time-2h at RT
Sample Volume-10 µL
Shipping Conditions-Shipped in Blue Ice
Target-STAT6
Target Class-Phosphoproteins
Target Species-Human
Technology-Alpha
Therapeutic Area-Inflammation Oncology
Unit Size-10,000 assay points