PAb Anti-phospho PKC substrate-Eu cryptate enables the rapid detection of phosphorylation of peptides or proteins on Ser in a biochemical kinase assay format.
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  • No-wash No-wash
  • Ease-of-use Ease-of-use
  • High affinity High affinity
PAb Anti-phospho PKC substrate-Eu cryptate enables the rapid detection of phosphorylation of peptides or proteins on Ser in a biochemical kinase assay format.
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Overview

The Anti-phospho PKA-Eu cryptate is a labelled polyclonal antibody. It recognizes phosphorylated serine with arginine or lysine residues in the -2 and +2 positions and a hydrophobic residue in the +1 position. It does not cross-react with non-phosphorylated serine residue or phosphorylated serine in a different configuration, nor does it recognize phospho threonine. 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. HTRF allows to perform assay at physiological ATP concentrations.

Benefits

  • NO LIMITATION IN ATP CONCENTRATIONS
  • ANY KIND OF SUBSTRATES
  • PHOSHO-(SER) SPECIFIC

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.
HTRF kinase assay principle

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 PKC substrate-Eu cryptate , prepared in a buffer containing EDTA to stop the enzymatic reaction.
HTRF kinase assay protocol
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Product Insert A-P PKC-Eu / 61P03KAE-61P03KAZ-61P03KAY

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