Hyper sensitive protein detection by Tandem-HTRF reveals Cyclin D1 dynamics in adult mouse

• DOI: 10.1038/srep15739
• Publication Date: 2015-10-27T10:38:52Z
• Authors: Alexandre Zampieri; Julien Champagne; Baptiste Auzemery; Ivanna Fuentes; Benjamin Maurel; Frédéric Bienvenu
• Keywords: Tandem

Abstract We present here a novel method for the semi-quantitative detection of low abundance proteins in solution that is both fast and simple. It based on Homogenous Time Resolved Förster Resonance Energy Transfer (HTRF), between lanthanide labeled donor antibody d2 or XL665 acceptor are raised against different epitopes same target. This approach we termed “Tandem-HTRF”, can specifically reveal rare polypeptides from only few microliters cellular lysate within one hour 384-well plate format. Using this sensitive approach, observed surprisingly core cell cycle regulator Cyclin D1 sustained fully developed adult organs harbors an unexpected expression pattern affected by environmental challenge. Thus our method, Tandem-HTRF offers promising way to investigate subtle variations dynamics sparse limited biological material.