Revvity, ASBF-TST5-A500, AlphaLISA SureFire Biotin-Free Human and Mouse Total STAT5 Detection Kit, 500 Assay Points

Overview
STAT5, comprising STAT5A and STAT5B, is activated by JAK kinases in response to cytokine signaling. Upon phosphorylation, STAT5 forms dimers and translocates to the nucleus, inducing transcription of genes involved in proliferation, differentiation, and survival. Overactive STAT5 drives cancers like leukemia and breast cancer, while its dysregulation contributes to autoimmune diseases by promoting excessive immune responses.
The AlphaLISA SureFire Biotin-Free Human and Mouse Total STAT5 Detection Kit is a sandwich immunoassay for the quantitative detection of total STAT5 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 Biotin Free kits are compatible with:

Cell and tissue lysates
Antibody modulators
Biotherapeutic antibodies
Biotin rich samples

AlphaLISA SureFire Biotin Free kits can be used for:

Cellular kinase assays
Receptor activation studies
High-throughput screening for preclinical studies
How it works
Total-AlphaLISA SureFire Biotin Free assay principle
The Total-AlphaLISA Surefire Biotin Free assay measures the expression level of a protein target in a cell lysate.
The Total-AlphaLISA Surefire assay uses two antibodies which recognize two different distal epitopes 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 targeted 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 protein present in the sample.

 

Total-AlphaLISA SureFire 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 Total-AlphaLISA SureFire 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.

Total-AlphaLISA SureFire one-plate assay protocol
Detection of Total target protein with AlphaLISA SureFire 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 quality.
Application-Cell Signaling
Automation Compatible-Yes
Brand-AlphaLISA SureFire Biotin-Free
Detection Modality-Alpha
Product Group-Kit
Protocol Time-2h at RT
Shipping Conditions-Shipped in Blue Ice
Target-STAT5
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-Alpha
Therapeutic Area-Autoimmunity Oncology
Unit Size-500 assay points