MAb Anti-phospho p53-Eu cryptate enables the rapid detection of p53 phosphorylation on Ser15 in a biochemical kinase assay format.
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  • No-wash
  • Ease-of-use
  • High affinity
MAb Anti-phospho p53-Eu cryptate enables the rapid detection of p53 phosphorylation on Ser15 in a biochemical kinase assay format.
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Overview

The Anti-phospho p53-Eu cryptate is a labeled monoclonal antibody enabling the rapid detection of p53 only when phosphorylated at Ser15, it does not cross-react with the corresponding non-phosphorylated sequence. 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-phospho p53-Eu cryptate , prepared in a buffer containing EDTA to stop the enzymatic reaction.
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