Revvity, 61P17KAZ, HTRF Anti-Phospho-MEK1/2 (Ser217/221) pAb Eu-Conjugate, 10,000 Assay Points

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.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
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.

 

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. 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-10,000 assay points