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Journal of Biomolecular Screening
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Article

High-Throughput Flow Cytometry-Based Assay to Identify Apoptosis-Inducing Proteins

Mamatha Sauermann1, Florian Hahne, Christian Schmidt, Meher Majety, Heiko Rosenfelder, Stephanie Bechtel, Wolfgang Huber, Annemarie Poustka, Dorit Arlt, Stefan Wiemann2*

1 Division of Molecular Genome Analysis, German Cancer Research Centre
2 German Cancer Research Center - DKFZ

* To whom correspondence should be addressed. E-mail: s.wiemann{at}dkfz.de.


  Abstract
After sequencing the human genome, the challenge ahead is to systematically analyze the functions and disease relation of the proteins encoded. Here the authors describe the application of a flow cytometry-based high-throughput assay to screen for apoptosis-activating proteins in transiently transfected cells. The assay is based on the detection of activated caspase-3 with a specific antibody, in cells overexpressing proteins tagged C- or N-terminally with yellow fluorescent protein. Fluorescence intensities are measured using a flow cytometer integrated with a high-throughput autosampler. The applicability of this screen has been tested in a pilot screen with 200 proteins. The candidate proteins were all verified in an independent microscopy-based nuclear fragmentation assay, finally resulting in the identification of 6 apoptosis inducers. (Journal of Biomolecular Screening XXXX:xx-xx)

First published on May 3, 2007, doi:10.1177/1087057107301271

Journal of Biomolecular Screening 2007;12:510.

A more recent version of this article appeared on June 1, 2007

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