How to measure intracellular pH using flow cytometry?
Posted April 2, 2019
RatioWorks™ BCFL, AM (21190) is a cell-permeable fluorogenic probe that can be used in flow cytometric assays to measure intracellular pH. Below is a brief protocol:
- Prepare cells for flow cytometry – for optimal performance flow cytometry requires between 2 x 105 to 1 x 106 cells
- Centrifuge to remove growth medium
- Add 1X 100 nM RatioWorks™ BCFL, AM working solution
- Incubate at 37 °C for 30 to 60 minutes
- Centrifuge to remove the dye loading solution
- Re-suspend cells in 1 mL Hank's Buffer with Hepes (HHBS).
- Analyze your sample with a flow cytometer using FITC channel