BCECF-AM is a cell membrane permeable compound widely used as a fluorescent indicator for intracellular pH. Storage: protect from light.

Fully treated cells show hydrogenosomes with an electron-dense deposit which aggregates to a variable extent.The staining is seen in the interior of hydrogenosomes in some instances. It is also observed by microscopy that the K⁺/H⁺ ionophor nigericin does not inhibit hydrogenosomal loading with BCECF-AM.
The pH-sensitive fluorescent dyes to measure cytosolic pH.
1.Prepare a 2 to 20 mM stock solution of BCECF-AM in DMSO.
2.Prepare a 5-50 µM BCECF-AM dye-loading solution in buffer solutions (HHBS or PBS).
3. Add 1000 µL/well (6-well plate),100 µL/well (96-well plate) or 25 µL/well (384-well plate) BCECF-AM dye-loading solution into the cell plate.
4. Incubate the dye-loading plate in a cell incubator for 30-60 minutes.
5. Wash and replace the dye-loading solution with buffers.
6. Run the pH assay by monitoring the fluorescence at E_x/E_m = 490/535 nm or 430/535 nm for ratio measurements.

西域 has not independently confirmed the accuracy of these methods. They are for reference only.

• . Scott DA, et al. Analysis of the uptake of the fluorescent marker 2',7'-bis-(2-carboxyethyl)-5(and-6)-carboxyfluorescein (BCECF) by hydrogenosomes in Trichomonas vaginalis. Eur J Cell Biol. 1998 Jun;76(2):139-45.  [Content Brief]

  . Clark Undem, et al. Endothelin-1 Augments Na+/H+ Exchange Activity in Murine Pulmonary Arterial Smooth Muscle Cells via Rho Kinase. PLoS One. 2012; 7(9): e46303.