HTRF PAb Anti-phospho MEK1/2-Eu cryptate, 500 Assay Points

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HTRF PAb Anti-phospho MEK1/2-Eu cryptate, 500 Assay Points

PAb Anti-phospho MEK1/2-Eu cryptate image

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.

For research use only. Not for use in diagnostic procedures.

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Feature	Specification
Application	Biochemical Enzymatic Assay

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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.

For research use only. Not for use in diagnostic procedures.

View product information

Product Variants

Unit Size: 500 Assay Points

Part #:

61P17KAE

List Price

USD 872.77

Unit Size: 10,000 Assay Points

Part #:

61P17KAZ

List Price

USD 4,888.00

Product information

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.

Specifications

Other Specifications

Application	Biochemical Enzymatic Assay
Brand	HTRF
Detection Modality	HTRF
Molecular Modification	Phosphorylation
Product Group	Fluorescent Reagent
Shipping Conditions	Shipped in Dry Ice
Target Class	Kinase
Technology	TR-FRET
Therapeutic Area	Cardiovascular Infectious Diseases Inflammation Metabolism/Diabetes NASH/Fibrosis Neuroscience Oncology & Inflammation Rare Diseases
Unit Size	500 Assay Points

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How it works

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.

HTRF PAb Anti-phospho MEK1.2-Eu cryptate 1.svg

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.

HTRF PAb Anti-phospho MEK1.2-Eu cryptate 2.svg

Assay validation

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