title: TR-FRET Assays for Endogenous Huntingtin Protein Level in Mouse Cells.
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).
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: Yijian
authors.initials: Y
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: Yuwei
authors.initials: Y
authors.lastname: Yao
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: Mingxing
authors.initials: M
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: Jiapeng
authors.initials: J
authors.lastname: Hou
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: Shenliang
authors.initials: S
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China Mass General Institute for Neurodegenerative Disease, Charlestown, MA, USA.
authors.forename: Boxun
authors.initials: B
authors.lastname: Lu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004195:Disease Models, Animal; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods; D000071058:Huntingtin Protein; D007118:Immunoassay / Q000379:methods*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism*
chemical_list: C506812:Htt protein, mouse; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins
keywords: Huntington's disease; TR-FRET; high-throughput technology; protein quantification