Revvity, 61P09KAE, HTRF Anti-Phospho-SAP/JNK (Thr183/Tyr184) mAb Eu-Conjugate, 500 Assay Points

Overview
The Anti-phospho SAP/JNK-Eu cryptate is a labeled monoclonal antibody enabling the rapid detection of SAP/JNK only when phosphorylated at Thr183 & Tyr185. It does not cross-react with phosphorylated forms of p44/42 or p38 MAPK. It does not detect the non-phosphorylated SAP/JNK. 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.
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 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-phosphoSAP/JNK-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