MAb Anti-phospho p53-Eu cryptate
Anti;phospho p53;Eu cryptate to detect phospho;p53;Ser15.
PAb Anti-phospho MEK1/2-Eu cryptate
PAb Anti-phospho MEK1/2-Eu cryptate enables the rapid detection of MEK1/2 phosphorylation on Ser 217/221 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 MEK1/2 substrate-Eu cryptate is a labeled polyclonal antibody enabling the detection of MEK1/2 when phosphorylated at Ser217/221. It recognizes singly phosphorylated MEK1/2 at Ser 217 but not MEK1/2 singly phosphorylated at Ser 221. It does not detect the non-phosphorylated MEK1/2. This allows the construction of a specific biochemical kinase assay. Any kind of substrate can be used, from small synthetic peptides to natural substrates, meaning high assay detection versatility. HTRF allows to perform assay at physiological ATP concentrations.
Benefits
- NO LIMITATION IN ATP CONCENTRATIONS
- ANY KIND OF SUBSTRATES
- PHOSHO-MEK1/2 (SER217/221) SPECIFIC
Assay principle
HTRF kinase assay use typically 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 PAb Anti-phospho MEK1/2-Eu cryptate , prepared in a buffer containing EDTA to stop the enzymatic reaction.
Working with natural ligand, development of COT assay
As shown here, the HTRF Kinase toolbox reagents can be used with any tagged peptide or protein substrates. The performances of each format were compared to P33 filter-binding assay in the study published by J. Jia (Anal. Biochem. 2006:305:268-276 ). In format 1, anti-GST-XL665 and PAb Anti-phospho MEK1/2-Eu cryptate were used to detect the whole GST-tagged protein, while in format 2, a small biotinylated MEK1 peptide was detected using Streptavidin-XL665 and the same phospho specific labeled antibody. Results are published in the table below
Apparent ATP Km values determined by different COT assay formats
| Assay Format | Apparent ATP Km(µM) |
|---|---|
| COT/MBP protein direct33P | 384 ± 55 |
| COT/MEK protein direct HTRF | 279 ± 40 |
| COT/MEK peptide direct HTRF | 268 ± 21 |
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 MEK1/2-Eu / 61P17KAE-61P17KAZ-61P17KAY
61P17KAE-61P17KAZ-61P17KAY - 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 MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet (DEU) A-P MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet (ELL) A-P MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet (FRA-FR) A-P MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet (ITA) A-P MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet (SPA) A-P MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet (ENG-GB) A-P MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet (ENG-US) A-P MEK1/2-Eu / 61P17KAE
61P17KAE - Safety Data Sheet
Safety Data Sheet A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet (DEU) A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet (ELL) A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet (FRA-FR) A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet (ITA) A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet (SPA) A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet (ENG-GB) A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet (ENG-US) A-P MEK1/2-Eu / 61P17KAY
61P17KAY - Safety Data Sheet
Safety Data Sheet A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
Safety Data Sheet (DEU) A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
Safety Data Sheet (ELL) A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
Safety Data Sheet (FRA-FR) A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
Safety Data Sheet (ITA) A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
Safety Data Sheet (SPA) A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
Safety Data Sheet (ENG-GB) A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
Safety Data Sheet (ENG-US) A-P MEK1/2-Eu / 61P17KAZ
61P17KAZ - Safety Data Sheet
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