Heinen_Transcription factor staining with retention of fluorescent proteins

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. 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.