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AlphaLISA SureFire Ultra Human and Mouse Phospho-AKT1/2/3 (Thr450) Detection Kit, 500 Assay Points

The AlphaLISA™ SureFire® Ultra™ Human and Mouse Phospho-AKT 1/2/3 (Thr450) assay is a sandwich immunoassay for quantitative detection of phospho-AKT 1/2/3 (Thr450) in cellular lysates using Alpha Technology.

Feature Specification
Application Cell Signaling
Sample Volume 10 µL

The AlphaLISA™ SureFire® Ultra™ Human and Mouse Phospho-AKT 1/2/3 (Thr450) assay is a sandwich immunoassay for quantitative detection of phospho-AKT 1/2/3 (Thr450) in cellular lysates using Alpha Technology.

Product variants
Unit Size: 500 assay points
Part #:
ALSU-PAKT-C500
List price
USD 2,392.92
Your online price:
Unit Size: 10,000 assay points
Part #:
ALSU-PAKT-C10K
List price
USD 14,400.00
Your online price:
Unit Size: 50,000 assay points
Part #:
ALSU-PAKT-C50K
List price
USD 46,000.00
Your online price:
Unit Size: 100 assay points
Part #:
ALSU-PAKT-C-HV
List price
USD 708.33
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

Protein kinase B (PKB) also known as AKT1 AKT2 and AKT3 encode the RAC alpha, beta, and gamma serine/threonine protein kinases respectively. AKT1 is involved in cellular survival pathways, by inhibiting apoptotic process. It is also able to induce protein synthesis pathways, and is therefore a key signaling protein in the cellular pathways that lead to skeletal muscle hypertrophy and general tissue growth. Akt2 is an important signaling molecule in the insulin signaling pathway. AKT3 appears to be predominantly expressed in the brain.

The AlphaLISA SureFire Ultra Human and Mouse Phospho-AKT 1/2/3 (Thr450) Detection Kit is a sandwich immunoassay for the quantitative detection of phospho-AKT 1/2/3 (Thr450) 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

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.

Specifications

Application
Cell Signaling
Automation Compatible
Yes
Brand
AlphaLISA SureFire Ultra
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
AKT 1/2/3
Target Class
Phosphoproteins
Target Species
Human
Mouse
Technology
Alpha
Therapeutic Area
Cardiovascular
Oncology
Unit Size
500 assay points

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