Fura Red

Guide^[2] (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Labeling of Cells:
1. Resuspend the cells in Hanks' Balanced Salt Solution (HBSS), add 1 μM Fura Red AM, supplement with 0.01% Pluronic F127 unless otherwise stated, and incubate in a 37°C water bath for 30 minutes.
2. Wash the cells in HEPES-buffered salt solution (HBSS) containing 1 mmol/L calcium chloride, 0.5 mmol/L magnesium chloride, 0.1% bovine serum albumin, and 10 mmol/L HEPES, then resuspend them in the same HEPES-buffered salt solution supplemented with the vital dye SYTOX Green.
3. Equilibrate the cells in a 37°C water bath for at least 10 minutes.
4. Analyze the samples on a flow cytometer, fluorescence microscope, or fluorescence microplate reader. Ratiometric analysis of Fura Red is performed by excitation with a violet laser (406 nm) and a green laser (502 nm).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.