Revvity, 61P01KAE, HTRF Anti-Phospho-PKA substrate pAb Eu-Conjugate, 500 Assay Points

Overview
The Anti-phospho PKA-Eu cryptate is a labeled polyclonal antibody enabling the detection of peptides and proteins containing a phosphorylated Ser/Thr residue with Arginine in the 3 position. It does not detect the non-phosphorylated PKA. This allows the construction of a highly specific biochemical kinase assay. Any kind of substrates can be used, from small synthetic peptides, to natural substrates for a high assay detection versatility. As HTRF tolerates extremely high ATP concentrations, an assay can be performed 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 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 assay 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 PKA substrate-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-500 assay points