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Experiment Overview

Repository ID: FR-FCM-ZZ9P Experiment name: Heinen_Transcription factor staining with retention of fluorescent proteins MIFlowCyt score: 49.71%
Primary researcher: André Heinen PI/manager: Florian Kurschus Uploaded by: Florian Kurschus
Experiment dates: 2012-04-25 - 2013-12-11 Dataset uploaded: Jan 2014 Last updated: Feb 2014
Keywords: [flow cytometry] [fixation] [EYFP] [RFP] [EGFP] [FoxP3] [RORγt] [T-bet] [fate mapping] [formaldehyde] [mouse T cells.] Manuscripts: [24616430] Cytalogo
Organizations: University Medical Center of the Johannes Gutenberg-University Mainz, Institute for Molecular Medicine, Mainz, (Germany)
Purpose: To develop a method which would allow simultaneous staining of nuclear TFs such as FoxP3, ROR?t or T-bet in cells, which express a fluorescent reporter protein.
Conclusion: We elaborated a simple and reliable protocol, which allows efficient transcription factor and cytokine staining while retaining fluorescent proteins inside fixed cells.
Comments: None
Funding: Not disclosed
Quality control: Internal quality control procedures were used to assess instrument parameters and assure accurate results. Daily calibration of the flow cytometer was performed using the CST module. Before use, antibodies where titrated to obtain the optimal staining. Experiments were repeated up to 3 times, also by different experimentators.
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