TR-FRET Assays for Endogenous Huntingtin Protein Level in Mouse Cells

Article type: Short Communication

Authors: Liang, Yijian | Yao, Yuwei | Lu, Mingxing | Hou, Jiapeng | Yu, Shenliang | Lu, Boxun^;

Affiliations: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China | Mass General Institute for Neurodegenerative Disease, Charlestown, MA, USA

Note: [] These authors contributed equally.

Note: [] These authors contributed equally.

Note: [] These authors contributed equally.

Note: [] Correspondence to: Boxun Lu, 220 Handan Road, Fudan University. E-mail: [email protected]

Abstract: High-throughput measurement of huntingtin (Htt) levels is useful for Huntington's disease research. For example, identification of genetic or chemical modifiers that reduce Htt levels by high-throughput screening provides promising strategy for HD drug discovery. In the human cells, high-throughput measurement of Htt levels has been established based on the Time Resolved-Fluorescence Resonance Energy Transfer (TR-FRET) technology, using the 2B7/MW1 antibody pair. Unfortunately, application of this assay in the mouse cells has been problematic due to discrepancies between TR-FRET signals and Western-blots, possibly caused by non-specific antibody binding. Here we report TR-FRET assays that are able to detect endogenous Htt levels of the mouse striatal cell line (STHdh).

Keywords: Huntington's disease, high-throughput technology, protein quantification, TR-FRET

DOI: 10.3233/JHD-140104

Journal: Journal of Huntington's Disease, vol. 3, no. 3, pp. 253-259, 2014