Dojindo,BCECF-AM/1/B262,BCECF 是使用细胞内 pH 探针

General Protocol (for Human Neutrophil)*Reagents:– 1 mM BCECF-AM/DMSO solution (1 mg BCECF in 1.45 ml DMSO)– HEPES buffer saline (20 mM HEPES, 153 mM NaCl, 5 mM KCl, 5 mM glucose, pH 7.4)

Protocol:1. Suspend cells in HEPES buffer solution to prepare 4×107 cells per ml.2. Add 1 mM BCECF-AM/DMSO solution to the cell suspension to prepare 3 μM BCECF-AM (1/300 vol of cell suspension) as the final concentration.3. Incubate the cell suspension at 37ºC for 30 minutes.4. Wash the cells 3 times with HEPES buffer saline and then prepare 3×10⁶ cells per ml of the cell suspension.5. Determine the fluorescence intensity using a fluorescence microscope or a confocal laser microscope coupled with an image analyzer.

* Cell staining conditions depend on cell type, so it is necessary to optimize the conditions for each experiment

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