Revvity, ALSU-TGS3B-A50K, AlphaLISA SureFire Ultra Human and Mouse Total GSK3β Detection Kit, 50,000 Assay Points

Overview
Glycogen synthase kinase-3 beta (GSK3β) is active in numerous central intracellular signaling pathways and regulates important cellular processes, from proliferation and immune response to glucose regulation and apoptosis. GSK3 kinase phosphorylation inhibits the activity of downstream targets such as β-catenin. GSK3 is regulated by cellular localization and by protein complex formation in the insulin pathway. Insulin activates PI3K, which phosphorylates AKT and in turn, activated AKT phosphorylates GSK3α on Ser21 and GSK3β on Ser9, resulting in GSK3 inactivation and inducing activation of glycogen synthase by dephosphorylation. GSK3 is considered a versatile target for various disease research applications, including oncology, neurological disorders, and diabetes.
The AlphaLISA SureFire Human and Mouse Total GSK3β Detection Kit is a sandwich immunoassay for the quantitative detection of total GSK3β 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

Alpha SureFire kits can be used for

Cellular kinase assays
Receptor activation studies
Screening
How it works
Total-AlphaLISA SureFire Ultra assay principle

The Total-AlphaLISA SureFire Ultra assay measures the expression level of a protein target in a cell lysate.

The Total-AlphaLISA SureFire Ultra 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 streptavidin to capture one of the detection antibodies, which is biotinylated. 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 Ultra 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 Ultra 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 Ultra one-plate assay protocol

Detection of Total target protein with AlphaLISA SureFire Ultra 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 AlphaLISA SureFire Ultra quality.
Application-Cell Signaling
Automation Compatible-Yes
Brand-AlphaLISA SureFire Ultra
Detection Modality-Alpha
Lysis Buffer Compatibility-Lysis Buffer
Molecular Modification-Total
Product Group-Kit
Sample Volume-10 µL
Shipping Conditions-Shipped in Blue Ice
Target-GSK3β
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-Alpha
Therapeutic Area-Metabolic Neuroscience Oncology
Unit Size-50,000 assay points