A method for determining enzyme kinetic parameters (Km, Kcat) using HTRF...
MAb Anti-phospho AKT-Eu cryptate HTRF®
MAb Anti-phospho AKT-Eu cryptate enables the rapid detection of Akt phosphorylation on Ser473 in a biochemical kinase assay format.
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No-wash
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Ease-of-use
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High affinity
Overview
The Anti-phospho AKT-Eu cryptate is a labeled monoclonal antibody enabling the rapid detection of Akt only when phosphorylated at Ser473. This allows the construction of a highly specific biochemical kinase assay. Any kind of substrate can be used, from small synthetic peptides to natural substrates, meaning high assay detection versatility. As HTRF tolerates extremely high ATP concentrations, an assay can be performed at physiological ATP concentrations.
Benefits
- NO LIMITATION IN ATP CONCENTRATIONS
- ANY KIND OF SUBSTRATES
- PHOSPHO-AKT (SER473) SPECIFIC
Assay principle
HTRF kinase assays typically use a combination of a Europium-cryptate labeled anti-phosphoresidue antibody, SA-XL665 and a biotinylated* substrate. The signal is proportional to the concentration of phospho-residues. *Alternatively, other tags such as 6HIS, GST, c-myc, DNP, or FLAG may be used to label the kinase substrate.
Assay protocol
HTRF kinase assays have two basic phases: 1. Enzymatic step: biotinylated substrate is incubated with the kinase of interest and with the compounds in presence of co-factors. The reaction starts with the addition of ATP. 2. Detection step: The phosphorylated biotinylated substrate is then detected by the addition of Streptavidin-XL665 and MAb Anti-phospho AKT-Eu cryptate , prepared in a buffer containing EDTA to stop the enzymatic reaction.
Open R&D: Sanofi Access Platform
In collaboration with Sanofi - Scientific Presentations
Experience the freedom of building your own assay
HTRF kinase activity toolbox reagents - Flyers
Product Insert A-P AKT-Eu / 61P15KAE-61P15KAZ-61P15KAY
61P15KAE-61P15KAZ-61P15KAY - Product Insert
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
The essential white paper to expand your knowledge on kinases
Learn about cutting edge kinase research in this free white paper - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Revolutionize your kinases investigations with HTRF
Watch this video to learn more about kinases investigations - Videos
Easy method for kinetic binding determination
How to revolutionize your kinetic binding demonstration with HTRF kinase binding platform assays - Application Notes
Safety Data Sheet A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet (DEU) A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet (ELL) A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet (FRA-FR) A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet (ITA) A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet (SPA) A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet (ENG-GB) A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet (ENG-US) A-P AKT-Eu / 61P15KAE
61P15KAE - Safety Data Sheet
Safety Data Sheet A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet (ELL) A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet (FRA-FR) A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet (ITA) A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet (SPA) A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet (ENG-GB) A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet (DEU) A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet (ENG-US) A-P AKT-Eu / 61P15KAY
61P15KAY - Safety Data Sheet
Safety Data Sheet A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Safety Data Sheet (DEU) A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Safety Data Sheet (ELL) A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Safety Data Sheet (FRA-FR) A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Safety Data Sheet (ITA) A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Safety Data Sheet (SPA) A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Safety Data Sheet (ENG-GB) A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Safety Data Sheet (ENG-US) A-P AKT-Eu / 61P15KAZ
61P15KAZ - Safety Data Sheet
Plate Reader Requirement
Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.
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