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

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

SUBMITTER: Zampieri A

PROVIDER: S-EPMC4622077 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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

Zampieri Alexandre A Champagne Julien J Auzemery Baptiste B Fuentes Ivanna I Maurel Benjamin B Bienvenu Frédéric F

Scientific reports 20151027

We present here a novel method for the semi-quantitative detection of low abundance proteins in solution that is both fast and simple. It is based on Homogenous Time Resolved Förster Resonance Energy Transfer (HTRF), between a lanthanide labeled donor antibody and a d2 or XL665 labeled acceptor antibody that are both raised against different epitopes of the same target. This novel approach we termed "Tandem-HTRF", can specifically reveal rare polypeptides from only a few microliters of cellular ...[more]

PMID: 26503526

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