How to measure intracellular pH using flow cytometry?

Q: How to measure intracellular pH using flow cytometry?

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

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RatioWorks&trade; 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 &ndash; 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&trade; BCFL, AM working solution
Incubate at 37 &deg;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

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