Phospho-AKT3 (Ser473) cellular kit
Simple, all-in-one kit for robust detection of Phoshpo-ATK & Total ATK
Phospho-AKT1/2/3 (Thr308) cellular kit HTRF®
This HTRF kit is designed to monitor pan AKT phosphorylation on Thr308 as a readout of the activated Pi3K pathway.
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Ease-of-use
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High sensitivity
Overview
The Phospho-AKT (Thr308) kit is designed for robust quantification of AKT modulation, phosphorylated on Thr308 across all isoforms (AKT1/2/3). 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.
Benefits
- VALIDATED ON MULTIPLE CELL TYPES
- SPECIFICITY
Phospho-AKT (Thr308) assay principle
The Phospho-AKT (Thr308) assay measures AKT when phosphorylated at Thr308. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-AKT (Thr308) 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-AKT (Thr308) 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-AKT (Thr308) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-AKT (Thr308) 1-plate assay protocol
Detection of Phosphorylated AKT (Thr308) 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.
Phospho-AKT (Thr308) IGF dose-response on CHO-M1 cells
Results obtained on CHO cells plated using 2 cell densities (25,000 - 50,000 cells per well). After being starved for 24 hours, cells were stimulated for 10 minutes with various concentrations of IGF. Phosphorylated AKT on Thr308 was measured after 3.5 hours using the two-plate assay protocol.
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.
Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
Open R&D: Sanofi Access Platform
In collaboration with Sanofi - Scientific Presentations
Cell Signaling: Biomarkers, Phospho- & total-protein Assays - Flyers
HTRF cellular phospho-protein assays
Physiologically relevant results fo fast flowing research - Flyers
Biomarker and Cell Signaling Assays for Fibrosis and NASH
HTRF and Alpha solutions for NASH - Flyers
HTRF products and supporting scientific content to investifgate GPCRs - Flyers
Cell Signaling: Biomarkers, Phospho- & total-protein assays - Flyers
HTRF assays for Oncology and Inflammation
Signaling in the Immune System - Brochures
In collaboration with Molecular Devices - Posters
PI3K/AKT/mTor translational control pathway - Posters
Universal HTRF® phospho-protein platform: from 2D, 3D, primary cells to patient derived tumor cells
Analysis of a large panel of diverse biological samples and cellular models - Posters
From 2D, 3D cell cultures to xenografts: A smart HTRF platform to maximize anticancer drug discovery
One technology across all samples - Application Notes
HTRF phospho assays reveal subtle drug induced effects in tumor-xenografts
Tumor xenograft analysis: HTRF versus Western blot - Application Notes
HTRF cell-based phospho-protein data normalization
Valuable guidelines for efficiently analyzing and interpreting results - Application Notes
HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers
Increased flexibility of phospho-assays - Application Notes
HTRF and Tag-lite technologies in multidimensional cell-based assays for GPCR investigations
16 GPCR challenges, 1 solution - Posters
Multi-tissue cellular modeling and anlysis of insulin signaling - Posters
Best practices for analyzing brain samples with HTRF® phospho assays for neurosciences
Insider Tips for successful sample treatment - Technical Notes
HTRF Alpha-tubulin Housekeeping kit
Properly interpret your compound effect - Application Notes
Understanding GPCRs is the key to improved DD
In collaboration with GEN - Guides
Optimize your HTRF cell signaling assays on tissues
HTRF and WB compatible guidelines - Technical Notes
Key guidelines to successful cell signaling experiments
Mastering the art of cell signaling assays optimization - Guides
HTRF® cell signaling platform combined with iCell® Hepatocytes
A solution for phospho-protein analysis in metabolic disorders - Posters
HTRF phospho-assays reveal subtle drug-induced effects
Detailed protocol and direct comparison with WB - Posters
A single technology for 2D cells, 3D cells, and xenograft models - Posters
Best practices for analyzing tumor xenografts with HTRF phospho assays
Protocol for tumor xenograft analysis with HTRF - Technical Notes
How to run a cell based phospho HTRF assay
What to expect at the bench - Videos
Unleash the potential of your phosphorylation research with HTRF
Unmatched ease of use, sensitivity and specificity assays - Videos
Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening
In collaboration with Bayer - Scientific Presentations
STING HTRF offer to bridge innate and adaptive immunity
cGAS-STING signaling pathway from A to Z - Brochures
Product Insert AKT P-T308 Kit / 64AKTPEG-64AKTPEH
64AKTPEG-64AKTPEH - Product Insert
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Unleash the potential of your phosphorylation research with HTRF
A fun video introducing you to phosphorylation assays with HTRF - Videos
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Videos
Molecular basis of neuroinflammation and neurodegeneration diseases
The essential guide for extending your knowledge on the molecular mechanisms of neurodegenerative diseases - Guides
Neurodegeneration and its main related diseases
Discover this infographic design on neurodegenerative diseases - Infographics
On-demand webinar: Linking Neuroinflammation and Neurodegeneration
New insight into neuroinflammation research - Videos
Assays for autoimmunity research
Advance your research on autoimmune diseases - Flyers
Assays for neurosciences research
Advance your research on neurodegenerative diseases - Flyers
Inhibition of FGFR Signaling Pathways Studied in Cancer Cell Lines Using HTRF
The knowledge and data necessary to understand the inhibition value of FGFR receptors involved in human cancers. HTRF technology enables the monitoring of the activity of the AZD4547 inhibitor on cell signal transduction in 3 cancer cell lines. - Application Notes
Advance your research on Fibrosis
Kits and reagents for Fibrosis research - Flyers
Signaling pathways in liquid tumors
This litterature review highlights the signaling pathways and give examples of how HTRF® KinEASE™ and AlphaLISA® SureFire® assays can be used in the development of novel therapies for leukemia treatment - Application Notes
Guidelines for Cell Culture and Lysis in Different Formats Prior to HTRF Detection
Seeding and lysing recommendations for a number of cell culture vessels. - Technical Notes
Assessment of drug efficacy and toxicity by combining innovative technologies
Combination of AlphaLISA®, HTRF®, or AlphaLISA® SureFire® Ultra™ immunoassays with the ATPlite™ 1step cell viability assay - Application Notes
Methodological Aspects of Homogeneous Time-Resolved Fluorescence (HTRF)
Learn how to reduce time and sample consumption - Application Notes
Safety Data Sheet AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet (DEU) AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet (ELL) AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet (ITA) AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet (SPA) AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) AKT P-T308 Kit / 64AKTPEG
64AKTPEG - Safety Data Sheet
Safety Data Sheet AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Safety Data Sheet (DEU) AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Safety Data Sheet (ELL) AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Safety Data Sheet (ITA) AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Safety Data Sheet (SPA) AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Safety Data Sheet (ENG-US) AKT P-T308 Kit / 64AKTPEH
64AKTPEH - Safety Data Sheet
Plate Reader Requirement
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