Revvity, 61Y20KLB, HTRF Anti-PY20 mAb Eu-Conjugate, 20,000 Assay Points

Overview
PY20-Eu cryptate is a labeled mouse monoclonal IgG2b. It shows to react specifically with phosphorylated tyrosine, both as free amino acid or when conjugated to carriers. No cross-reactivity has been observed with non-phosphorylated tyrosine, phosphothreonine, phosphoserine, AMP or ATP. This allows the construction of a highly specific biochemical Tyrosine 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.
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 Tyrosine kinase assay use typically a combination of a Europium-cryptate labeled anti-phosphotyrosine 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 PY20-Eu cryptate prepared in a buffer containing EDTA to stop the enzymatic reaction. Application-Biochemical Enzymatic Assay
Brand-HTRF
Detection Modality-HTRF
Product Group-Fluorescent Reagent
Shipping Conditions-Shipped Ambient
Target Class-Kinases
Technology-TR-FRET
Therapeutic Area-Cardiovascular Infectious Diseases Inflammation Metabolism/Diabetes NASH/Fibrosis Neuroscience Oncology & Inflammation Rare Diseases
Unit Size-20,000 assay points