HTRF Human Phospho-AKT2 (Ser473) Detection Kit, 500 Assay Points

Revvity Sites Globally

Select your location.

*e-commerce not available for this region.

Australia

Austria

Belgium

Brazil *

Canada

China *

Denmark

Finland

France

Germany

Hong Kong *

India *

Ireland

Italy

Japan *

Luxembourg

Mexico *

Netherlands

Norway

Philippines *

Republic of Korea *

Singapore *

Spain

Sweden

Switzerland

Thailand *

United Kingdom

United States

HTRF Human Phospho-AKT2 (Ser473) Detection Kit, 500 Assay Points

SDS

COAs

IFUs, Manuals & more

This HTRF kit is designed for robust cell-based quantification of AKT2 modulation, phosphorylated on Ser473.

Product information

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).

SDS

COAs

IFUs, Manuals & more

Product Variants

partnumber

Part number: 63ADK080PEG

Save to favorites

Unit Size: 500 Assay Points

List price: USD 2,147.00

Your price: Login to view

USD 2,147.00

USD 2,147.00 /each

partnumber

Part number: 63ADK080PEH

Save to favorites

Unit Size: 10,000 Assay Points

List price: USD 12,490.00

Your price: Login to view

USD 0.00

USD 12,490.00 /each

Product information

Overview

The Phospho-AKT2 (Ser473) kit is designed for the robust quantification of AKT2 modulation, phosphorylated on Ser473. Also known as protein kinase B (PKB), AKT is an oncogene playing a key role in controlling apoptosis, cell proliferation, transcription, cell migration, and glucose metabolism. AKT2 in particular plays a critical role in the insulin pathway, and thus has important applications in diabetes and metabolic research.

Specifications

Other Specifications

Assay Points	500
Assay Technology	HTRF
Brand	HTRF
Product Group	Kit
Quantity	1
Shipping Conditions	Shipped in Dry Ice
Therapeutic Area	Metabolism/Diabetes NASH/Fibrosis Oncology & Inflammation
Unit Size	500 Assay Points

Video gallery

How it works

Phospho-AKT2 (Ser473) assay principle

The Phospho-AKT2 (Ser473) assay measures AKT2 when phosphorylated at Ser473. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-AKT2 (Ser473) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the proteins phosphorylation state under a no-wash assay format.

Phospho-AKT2 (Ser473) 2-plate assay protocol

The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Phospho-AKT2 (Ser473) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

2biomarkers-how-it-works-assay-protocol-alpha-tubulin-63adk072peg-63adk072peh.svg

Phospho-AKT2 (Ser473) 1-plate assay protocol

Detection of Phosphorylated AKT2 (Ser473) with HTRF reagents can be performed in a single plate used for culturing, stimulation and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.

2phospho-how-it-works-total-btk-1-plate-assay-protocol.svg

Assay validation

Phospho AKT2 assay: Inhibitory effect of blocking peptide

SH-SY5Y cells were stimulated with human IGF-1 for 15 min. After stimulation, media was removed and cells were lysed with lysis buffer 1X for 30 min at RT under gentle shaking. 14 µL of lysates were transferred into 384-well sv white microplates and 2 µL of different concentration of blocking peptide specific for AKT2 or was added before adding 4 µL of the HTRF phospho-AKT2 (Ser473) detection reagents. The HTRF signal was recorded after an overnight incubation at room temperature.

Simplified pathway

AKT simplified pathway

AKT (or protein kinase B) plays a key role in controlling survival and apoptosis. This serine/threonine protein kinase is regulated by insulin and various growth and survival factors, to work in a wortmannin-sensitive pathway involving PI 3 kinase. When the Pleckstrin Homology (PH) domain of AKT binds to phosphoinositides, AKT can be phosphorylated by two different kinases, PDK1 at threonine 308 and mTORC2 (mammalian target of rapamycin) at serine 473, which switch on AKT activation.