Revvity, 61UBIKLA, HTRF Ubiquitin Eu-Conjugate, 5,000 Assay Points

Overview
Ubiquitination plays a key role in regulating protein degradation, and is carried out by the conjugation of ubiquitin to target proteins through the sequential activity of three enzymes: E1, E2 and E3. Poly-ubiquitinated proteins are then subject to degradation by proteasomes.
The ubiquitin pathway is associated with many pathologies such as cancer, muscle atrophy, inflammation, viral infection, and metabolic and neurodegenerative disorders. The Ubiquitin-Eu cryptate allows flexible assay development assessing each step in the ubiquitin pathway.
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 Ubiquitin assays use ubiquitin labeled with Europium cryptate. Upon the addition of XL665 conjugate, the HTRF signal is proportional to the level of ubiquitination, and remains stable for more than 24 hours. An example of a p53-polyubiquitination assay is shown here.

Assay protocol
HTRF ubiquitin assays have two basic phases: the enzymatic step, followed by the HTRF detection step. 1. Enzymatic step: The target protein is incubated with the ubiquitin enzymes (E2 & E3) and Eu3+-labeled ubiquitin. The reaction starts with the addition of E1 enzyme. 2. Detection step: The ubiquitinated target protein is then detected by the addition of an XL665-labeled reagent that contains EDTA to stop the enzymatic reaction. Application-Protein-Protein Interaction
Brand-HTRF
Detection Modality-HTRF
Product Group-Fluorescent Reagent
Shipping Conditions-Shipped Ambient
Target Class-Binding Assay
Technology-TR-FRET
Unit Size-5,000 assay points