Skip to main content
Menu

AlphaLISA SureFire Ultra Human Phospho-TIE2 (Tyr992) Detection Kit, 100 Assay Points

The AlphaLISA™ SureFire® Ultra™ Human Phospho-TIE2 (Tyr992) assay is a sandwich immunoassay for quantitative detection of phospho-TIE2 in cellular lysates using Alpha technology.

Feature Specification
Application Cell Signaling
Sample Volume 30 µL

The AlphaLISA™ SureFire® Ultra™ Human Phospho-TIE2 (Tyr992) assay is a sandwich immunoassay for quantitative detection of phospho-TIE2 in cellular lysates using Alpha technology.

Product variants
Unit Size: 100 assay points
Part #:
ALSU-PTIE2-A-HV
List price
USD 708.33
Your online price:
Unit Size: 500 assay points
Part #:
ALSU-PTIE2-A500
List price
USD 2,392.92
Your online price:
Unit Size: 10,000 assay points
Part #:
ALSU-PTIE2-A10K
List price
USD 14,400.00
Your online price:
Unit Size: 50,000 assay points
Part #:
ALSU-PTIE2-A50K
List price
USD 46,000.00
Your online price:
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).

Overview

The TEK receptor tyrosine kinase (TIE2), encoded by the TEK gene in humans, is primarily expressed on endothelial cells. Upon binding with angiopoietin-1 (Angpt1), TIE2 induces angiogenesis and supports endothelial cell survival. This activation leads to downstream signaling pathways, including MEK/ERK, IKKb/NFkB, and AKT. Consequently, TIE2 contributes to vascular stability, cellular proliferation, inflammation, and cell survival. Elevated levels of Ang1 and/or Ang2 have been observed in patients with various cancers.

The AlphaLISA SureFire Ultra Human Phospho-TIE2 (Tyr992) Detection Kit is a sandwich immunoassay for the quantitative detection of phospho-TIE2 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
  • High-throughput drug 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.

The referenced media source is missing and needs to be re-embedded.

 

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.

The referenced media source is missing and needs to be re-embedded.

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.

The referenced media source is missing and needs to be re-embedded.

Assay validation

Induction of TIE2 Phospho (Tyr992) in HUVEC cells

HUVEC cells were seeded in a 96-well culture plate (20,000 cells/well) in complete medium and incubated for 48 hours at 37°C, 5% CO2. The cells were further treated with increasing concentrations of Pervanadate for 30 minutes.

After treatment, the cells were lysed with 50 µL of lysis buffer for 10 minutes at RT with shaking at 350 rpm. TIE2 Phospho (Tyr992) and Total levels were evaluated using respective AlphaLISA SureFire Ultra assays. For the detection step, 10 µL of cell lysate (approximately 8,000 cells) were transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor Mix and incubated for 1 hour at RT.  Finally, 5 µL of Donor Mix was then added to each well and incubated for 1 hour at RT in the dark. The plate was read on an Envision Nexus™ using standard AlphaLISA settings.

Induction of TIE2 Phospho (Tyr992) in HUVEC cells

Specifications

Application
Cell Signaling
Automation Compatible
Yes
Brand
AlphaLISA SureFire Ultra
Detection Modality
Alpha
Host Species
Human
Lysis Buffer Compatibility
Lysis Buffer
Molecular Modification
Phosphorylation
Product Group
Kit
Sample Volume
30 µL
Shipping Conditions
Shipped in Blue Ice
Target
TIE2
Target Class
Phosphoproteins
Target Species
Human
Technology
Alpha
Therapeutic Area
Oncology
Unit Size
100 assay points

Video gallery

Resources

Are you looking for resources, click on the resource type to explore further.

1-1 of 1 Resources
Technical Note Icon
Technical Note
AlphaLISA assays for immune checkpoint detection and quantitation

Quantifying Immune Checkpoints: AlphaLISA Biomarker Detection Kits

This technical note focuses on the role of checkpoint molecules...

Loading...

Scroll Icon

Revvity AI Assistant Beta