MAb Anti-phosphop44/42-Eu cryptate

MAb Anti-phospho p44/42-Eu cryptate enables the rapid detection of p44/42 phosphorylation on Thr202 &Tyr204 in a biochemical kinase assay format.
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  • No-wash
  • Ease-of-use
  • High affinity
MAb Anti-phospho p44/42-Eu cryptate enables the rapid detection of p44/42 phosphorylation on Thr202 &Tyr204 in a biochemical kinase assay format.
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Overview

The Anti-phospho p44/42-Eu cryptate is a labeled monoclonal antibody enabling the rapid detection of p44/42 MAPK only when phosphorylated at Thr202 & Tyr204. It does not cross-react with phosphorylated forms of SAP/JNK or p38 MAPK. It does not detect the non-phosphorylated p44/42. 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.

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-phosphop44/42-Eu cryptate , prepared in a buffer containing EDTA to stop the enzymatic reaction.
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