BCECF-AM | Cytiva思拓凡-Whatman滤纸滤膜滤器

BCECF-AM;

BCECF-AM 是一种细胞膜穿透的荧光指示剂，可以测量细胞内的 pH 值。

BCECF-AM Chemical Structure

CAS No. : 117464-70-7

规格	价格	是否有货
1 mg	￥3700	询问价格货期

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生物活性	BCECF-AM is a cell membrane permeable compound widely used as a fluorescent indicator for intracellular pH.
体外研究 (In Vitro)	BCECF-AM is used to measure changes in basal pH_i and NHE activity induced by increasing concentrations of ET-1 (0.1-10 nM) in pulmonary arterial smooth muscle cell (PASMC)^[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量	820.70
Formula	C₈₀H₇₂O₃₈
CAS 号	117464-70-7
运输条件	Room temperature in continental US; may vary elsewhere.
储存方式	-20deg;C, protect from light *该产品在溶液状态不稳定，建议您现用现配，即刻使用。
参考文献	[1]. 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.

Cell Assay ^[1]	PASMCs are placed in a laminar flow cell chamber perfused with HBSS with pH adjusted to 7.4. pH_i is measured in cells incubated with the membrane permeant (acetoxymethyl ester) form of the pH-sensitive fluorescent dye BCECF-AM for 60 min at 37°C under an atmosphere of 20% O₂-5% CO₂. Cells are then washed with HBSS for 15 min at 37°C to remove extracellular dye and allow complete de-esterification of cytosolic dye. Ratiometric measurement of BCECF fluorescence is performed on a workstation consisting of a Nikon TSE 100 Ellipse inverted microscope with epi-fluorescence attachments. The light beam from a xenon arc lamp is filtered by interference filters at 490 and 440 nm, and focused onto the PASMCS under examination via a 20× fluorescence objective. Light emitted from the cell at 530 nm is returned through the objective and detected by an imaging camera. An electronic shutter is used to minimize photobleaching of dye. Protocols are executed and data collected on-line with InCyte software. pH_i is estimated from in situcalibration after each experiment. Cells are perfused with a solution containing (in mM): 105 KCl, 1 MgCl₂, 1.5 CaCl₂, 10 glucose, 20 HEPES-Tris and 0.01 nigericin to allow pH_i to equilibrate to external pH. A two point calibration is created from fluorescence measured as pH_i is adjusted with KOH from 6.5 to 7.5. Intracellular H⁺ ion concentration ([H⁺]i) is determined from pH_i using the formula: pH_i = −log ([H⁺]i). MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献	[1]. 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.

Cell Assay
^[1]

PASMCs are placed in a laminar flow cell chamber perfused with HBSS with pH adjusted to 7.4. pH_i is measured in cells incubated with the membrane permeant (acetoxymethyl ester) form of the pH-sensitive fluorescent dye BCECF-AM for 60 min at 37°C under an atmosphere of 20% O₂-5% CO₂. Cells are then washed with HBSS for 15 min at 37°C to remove extracellular dye and allow complete de-esterification of cytosolic dye. Ratiometric measurement of BCECF fluorescence is performed on a workstation consisting of a Nikon TSE 100 Ellipse inverted microscope with epi-fluorescence attachments. The light beam from a xenon arc lamp is filtered by interference filters at 490 and 440 nm, and focused onto the PASMCS under examination via a 20× fluorescence objective. Light emitted from the cell at 530 nm is returned through the objective and detected by an imaging camera. An electronic shutter is used to minimize photobleaching of dye. Protocols are executed and data collected on-line with InCyte software. pH_i is estimated from in situcalibration after each experiment. Cells are perfused with a solution containing (in mM): 105 KCl, 1 MgCl₂, 1.5 CaCl₂, 10 glucose, 20 HEPES-Tris and 0.01 nigericin to allow pH_i to equilibrate to external pH. A two point calibration is created from fluorescence measured as pH_i is adjusted with KOH from 6.5 to 7.5. Intracellular H⁺ ion concentration ([H⁺]i) is determined from pH_i using the formula: pH_i = −log ([H⁺]i).

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

参考文献

[1]. 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.