Related Experiment Video
Updated: Feb 10, 2026

Time-Resolved Fluorescence Anisotropy from Single Molecules for Characterizing Local Flexibility in Biomolecules
Published on: April 25, 2025
Miniaturizable homogenous time-resolved fluorescence assay for carboxypeptidase B activity
Marc Ferrer1, Paul Zuck, Garrett Kolodin
1Department of Automated Biotechnology, Merck Research Laboratories, North Wales, PA 19454, USA. marc_ferreralegre@merck.com
Abstract:
An epitope-unmasking, homogeneous time-resolved fluorescence (HTRF) assay has been developed for measuring carboxypeptidase B (CPB) activity in a miniaturized high-throughput screening format. The enzyme substrate (biotin-RYRGLMVGGVVR-OH) is cleaved by CPB at the C terminus, causing release of the C-terminal Arg residue. The product (biotin-RYRGLMVGGVV-OH) is recognized specifically by a monoclonal antibody (G2-10) which is labeled with Eu(3+)-cryptate ([Eu(3+)]G2-10 mAb), and the complex is detected by fluorescence resonance energy transfer using streptavidin labeled with allophycocyanin ([XL665]SA). The CPB HTRF assay is readily adapted from 96- to 1536-well format as a robust (Z(')>0.5) assay for high-throughput screening.
Related Concept Videos
Test for Homogeneity
Homogeneous Equilibria for Gaseous Reactions
For gas-phase reactions, the equilibrium constant may be expressed in terms of either the molar concentrations (Kc) or partial pressures (Kp) of the reactants and products. A relation between these two K values may be simply derived from the ideal gas equation and the definition of molarity. According to the ideal gas equation:
Tissue Homogenization and Cell Lysis
Mechanical methods of tissue homogenization
These methods rely on applying external physical force to disrupt...
Co-activators and Co-repressors
tRNA Activation
Activation Energy

