BCECF-AM

目录号: HY-101883: Data Sheet SDS COA 产品使用指南
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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 可以透过细胞膜扩散，细胞内酯酶裂解酯键，释放出 BCECF (HY-101882)。BCECF 允许测量生理 pH 值范围在 6.0-8.0 之间。激发波长: 490/440 nm; 发射波长: 535 nm。

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BCECF-AM Chemical Structure

CAS No. : 117464-70-7

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规格	价格	是否有货
205 μg (5 mM * 50 μL in DMSO)	￥1600	3 - 4 周
410 μg (5 mM * 100 μL in DMSO)	￥2620	3 - 4 周
820 μg (5 mM * 200 μL in DMSO)	￥4200	3 - 4 周

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BCECF-Acetoxyemnethy ester In-stock

MCE 顾客使用本产品发表的 7 篇科研文献

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生物活性
实验参考方法
纯度 & 产品资料
参考文献

生物活性	BCECF-AM is a cell membrane permeable compound widely used as a fluorescent indicator for intracellular pH. BCECF-AM could diffuse through the cell membrane and intracellular esterase cleave the ester bond releasing BCECF (HY-101882). BCECF allows measurements in the physiological pH range 6.0-8.0. Excitation ratio: 490/440 nm; Emission intensity: 535 nm.
体外研究 (In Vitro)	用于测量细胞溶质的 pH 敏感荧光染料pH^[3]^[4]。 1. 在 DMSO 中制备 5 mM BCECF-AM 原液。 2. 在 NMG 缓冲液 (10 mM HEPES、60 mM KCl、3 mM MgCl2，pH 6.8) 或 PBS 中制备 3-5 μM BCECF-AM 染料工作溶液。 3. 将 BCECF-AM 染料工作溶液 (3-5 µM) 加入细胞板中，37°C 避光孵育 30-60 分钟。 4. 用 PBS 洗涤细胞三次。 5. 通过监测 Ex/Em = 490/535 nm 或 440/535 nm 处的荧光来进行 pH 测定。标准曲线测定： 1. pH 校准缓冲液：130 mM KCl, 1 mM MgCl2, 15 mM HEPES, 15 mM MES (HY-D0858), 10 μM Nigericin sodium salt (HY-100381) 和 10 μM Valinomycin (HY-N6693) (使细胞内外 pH 达到平衡)。可用 NaOH 或者 HCl 调至 pH 为 6.5, 7.0, 7.5, 8.0 和 8.5。 2. 将染色后的细胞分别在 pH 6.5, 7.0, 7.5, 8.0 和 8.5 的校准缓冲液中孵育 10 分钟。 3. 检测各 pH 对应的 F490/F440 比值，以 pH 为横坐标，荧光比值为纵坐标，绘制标准曲线 (符合 S 形曲线，可通过 Henderson-Hasselbalch 方程拟合)。 4. 实验中测得的未知样品荧光比值 (R) 代入标准曲线，即可求出对应的 pH。 MCE has not independently confirmed the accuracy of these methods. They are for reference only. BCECF-AM 相关抗体:
分子量	820.70
Formula	C₄₀H₃₆O₁₉
CAS 号	117464-70-7
性状	液体
颜色	Orange to red
Emission (Em)	528
Excitation (Ex)	503
运输条件	Room temperature in continental US; may vary elsewhere.
储存方式	Solution, -20°C, protect from light, 2 years
纯度 & 产品资料	纯度: ≥99.0% 选择批次： Data Sheet (570 KB) SDS (393 KB) COA (305 KB) 产品使用指南 (1538 KB)
参考文献	[1]. 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] [2]. 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. [3]. Chen F, et al. CYP51A1 drives resistance to pH-dependent cell death in pancreatic cancer. Nat Commun. 2025 Mar 7;16(1):2278. [Content Brief] [4]. Pang X, et al. Weak acids produced during anaerobic respiration suppress both photosynthesis and aerobic respiration. Nat Commun. 2023 Jul 14;14(1):4207. [Content Brief]

Cell Assay ^[2]	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]. 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] [2]. 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. [3]. Chen F, et al. CYP51A1 drives resistance to pH-dependent cell death in pancreatic cancer. Nat Commun. 2025 Mar 7;16(1):2278. [Content Brief] [4]. Pang X, et al. Weak acids produced during anaerobic respiration suppress both photosynthesis and aerobic respiration. Nat Commun. 2023 Jul 14;14(1):4207. [Content Brief]

Cell Assay
^[2]

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]. 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]

[2]. 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.

[3]. Chen F, et al. CYP51A1 drives resistance to pH-dependent cell death in pancreatic cancer. Nat Commun. 2025 Mar 7;16(1):2278. [Content Brief]

[4]. Pang X, et al. Weak acids produced during anaerobic respiration suppress both photosynthesis and aerobic respiration. Nat Commun. 2023 Jul 14;14(1):4207. [Content Brief]

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质量		浓度		体积		分子量 *
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Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

浓度 _(start)	×	体积 _(start)	=	浓度 _(final)	×	体积 _(final)
	×		=		×
C1		V1		C2		V2

Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

BCECF-AM117464-70-7Fluorescent DyeInhibitorinhibitorinhibit

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BCECF-AM

Customer Review

Other Forms of BCECF-AM:

MCE 顾客使用本产品发表的 7 篇科研文献

BCECF-AM 相关产品

•同靶点产品:

•同靶点蛋白产品:

生物活性

实验参考方法

纯度 & 产品资料

参考文献

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BCECF-AM 相关抗体

MCE 顾客使用本产品发表的 7 篇科研文献

BCECF-AM 相关产品

•同靶点产品:

•同靶点蛋白产品:

生物活性

实验参考方法

纯度 & 产品资料

参考文献

BCECF-AM 相关抗体:

BCECF-AM 相关分类

摩尔计算器

稀释计算器

The molarity calculator equation

The dilution calculator equation

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