Revvity, ALSU-PERK-A50K, AlphaLISA SureFire Ultra Human and Mouse Phospho-ERK1/2 (Thr202/Tyr204) Detection Kit, 50,000 Assay Points

Overview
ERK1/2 (Extracellular Signal-Regulated Kinases 1 and 2), also known as MAPK3 and MAPK1 respectively, are crucial components of the MAPK/ERK signaling pathway in both humans and mice. These kinases play central roles in various cellular processes, including cell proliferation, differentiation, survival, and migration. Activation of ERK1/2 occurs through dual phosphorylation at specific threonine and tyrosine residues (Thr202/Tyr204 for human ERK1; Thr185/Tyr187 for human ERK2), which is essential for their kinase activity and subsequent regulation of downstream targets. The MAPK/ERK pathway is frequently dysregulated in various pathological conditions, particularly in cancer, developmental disorders, and neurodegenerative diseases. Aberrant activation of ERK1/2 can contribute to uncontrolled cell proliferation, resistance to apoptosis, and enhanced cell survival, often associated with tumor progression and therapy resistance. Conversely, insufficient ERK1/2 activation can lead to developmental abnormalities and impaired cellular responses to growth factors and other stimuli.
The AlphaLISA SureFire Ultra Human and Mouse Phospho-ERK1/2 (Thr202/Tyr204) Detection Kit is a sandwich immunoassay designed for the quantitative detection of phosphorylated ERK1/2 in cellular lysates from both human and mouse samples 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
Phospho-AlphaLISA SureFire Ultra assay principle
The Phospho-AlphaLISA SureFire Ultra 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 streptavidin to capture one of the detection antibodies, which is biotinylated. 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 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 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 one-plate assay protocol
Detection of Phosphorylated 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
Cellular or Signaling Pathway-MAPK
Detection Modality-Alpha
Lysis Buffer Compatibility-Lysis Buffer
Molecular Modification-Phosphorylation
Product Group-Kit
Sample Volume-10 µL
Shipping Conditions-Shipped in Blue Ice
Target-ERK1/2
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-Alpha
Therapeutic Area-Cardiovascular Central Nervous System Neuroinflammation
Unit Size-50,000 assay points