Phospho-Insulin Receptor-beta (Tyr1150/1151) cellular kit
All-in-one kit for robust detection of Phospho Insulin-Receptor beta
Phospho-AKT1/2/3 (Ser473) cellular kit HTRF®
This HTRF kit is designed to monitor pan Akt phosphorylation on Ser473 as a readout of Pi3K pathway activation.
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Ease-of-use
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High sensitivity
Overview
The Phospho-AKT (Ser473) kit is designed to robustly and efficiently monitor AKT phosphorylation on Ser473 across all isoforms, as a readout of Pi3K pathway activation. Also known as Protein Kinase B (PKB), AKT is an oncogene that controls apoptosis, cell proliferation, transcription, cell migration, and glucose metabolism.
Benefits
- VALIDATED ON MULTIPLE CELL TYPES
- SPECIFICITY
Phospho-AKT (Ser473) assay principle
The Phospho-AKT (Ser473) assay measures AKT when phosphorylated at Ser473. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-AKT (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-AKT (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-AKT (Ser473) assay HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-AKT (Ser473) assay 1-plate assay protocol
Detection of Phosphorylated AKT (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.
Phospho-AKT (Ser473) Assay IGF Dose-Response on HEK 293 cells
Results obtained on HEK cells plated using 3 cell densities (12,500 - 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 was measured after 3.5 hours using the two-plate assay protocol.
Total AKT used to control the phosphorylation status of AKT (Ser473).
HEK293 cells (100,000 cells/well) were activated with IGF-1 for 10 min, using the two-plate assay protocol of the Phospho-AKT (Ser473) and Total-AKT assays. As expected, results obtained show a dose-response increase of AKT phosphorylation upon IGF-1 stimulation, while the AKT expression level remains constant.
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
Cisbio lysis buffer compatibility
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 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
mechanism of action of EGFR inhibitors assessed in the pancreatic BXPC3 cell line - 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
A Homogeneous Assay to Quantify Endogenous AKT Phosphorylation in Human Umbilical Endothelial Cells
In collaboration with Biotek - 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
Product Insert AKT P-S473 Kit / 64AKSPET
64AKSPET - Product Insert
Homogeneous High Throughput Live Cell GPCR Functional and Surface Binding Assays
Molecular Devices - Posters
STING HTRF offer to bridge innate and adaptive immunity
cGAS-STING signaling pathway from A to Z - Brochures
Product Insert AKT P-S473 Kit / 64AKSPEG-64AKSPEH
64AKSPEG-64AKSPEH - Product Insert
HTRF Product Catalog 2020 July update
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
Safety Data Sheet AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet (DEU) AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet (ELL) AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet (ITA) AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet (SPA) AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) AKT P-S473 Kit / 64AKSPEG
64AKSPEG - Safety Data Sheet
Safety Data Sheet AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (DEU) AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (ELL) AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (ITA) AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (SPA) AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (ENG-US) AKT P-S473 Kit / 64AKSPEH
64AKSPEH - Safety Data Sheet
Safety Data Sheet (ELL) AKT P-S473 Kit / 64AKSPET
64AKSPET - Safety Data Sheet
Safety Data Sheet (FRA-FR) AKT P-S473 Kit / 64AKSPET
64AKSPET - Safety Data Sheet
Safety Data Sheet (ITA) AKT P-S473 Kit / 64AKSPET
64AKSPET - Safety Data Sheet
Safety Data Sheet (SPA) AKT P-S473 Kit / 64AKSPET
64AKSPET - Safety Data Sheet
Safety Data Sheet (ENG-GB) AKT P-S473 Kit / 64AKSPET
64AKSPET - Safety Data Sheet
Safety Data Sheet (ENG-US) AKT P-S473 Kit / 64AKSPET
64AKSPET - Safety Data Sheet
Safety Data Sheet (DEU) AKT P-S473 Kit / 64AKSPET
64AKSPET - Safety Data Sheet
Safety Data Sheet (DEU) AKT P-S473 Kit / 64AKSPEY
64AKSPEY - Safety Data Sheet
Safety Data Sheet (ELL) AKT P-S473 Kit / 64AKSPEY
64AKSPEY - Safety Data Sheet
Safety Data Sheet (FRA-FR) AKT P-S473 Kit / 64AKSPEY
64AKSPEY - Safety Data Sheet
Safety Data Sheet (ITA) AKT P-S473 Kit / 64AKSPEY
64AKSPEY - Safety Data Sheet
Safety Data Sheet (SPA) AKT P-S473 Kit / 64AKSPEY
64AKSPEY - Safety Data Sheet
Safety Data Sheet (ENG-GB) AKT P-S473 Kit / 64AKSPEY
64AKSPEY - Safety Data Sheet
Safety Data Sheet (ENG-US) AKT P-S473 Kit / 64AKSPEY
64AKSPEY - Safety Data Sheet
Plate Reader Requirement
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