1. Dye Reagents
  2. BCECF

BCECF (Synonyms: 2′,7′-Bis(2-carboxyethyl)-5(6)-carboxyfluorescein)

Cat. No.: HY-101882 Purity: ≥96.0%
Handling Instructions

BCECF is a pH-sensitive fluorescent dye and can be used to monitor cellular pH ratiometrically. BCECF allows measurements in the physiological pH range 6.0–8.0. Excitation ratio: 490/440 nm; Emission intensity: 535 nm.

For research use only. We do not sell to patients.

BCECF Chemical Structure

BCECF Chemical Structure

CAS No. : 85138-49-4

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1 mg USD 160 In-stock
Estimated Time of Arrival: December 31
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Based on 1 publication(s) in Google Scholar

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Description

BCECF is a pH-sensitive fluorescent dye and can be used to monitor cellular pH ratiometrically. BCECF allows measurements in the physiological pH range 6.0–8.0. Excitation ratio: 490/440 nm; Emission intensity: 535 nm.

In Vitro

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[1].
The pH-sensitive fluorescent dyes to measure cytosolic pH.
1.Prepare a 2 to 20 mM stock solution of BCECF in DMSO.
2.Prepare a 5-50 µM BCECF 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 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 Ex/Em = 490/535 nm or 430/535 nm for ratio measurements.

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

Molecular Weight

520.44

Formula

C₂₇H₂₀O₁₁

CAS No.
Shipping

Room temperature in continental US; may vary elsewhere.

Storage

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility
In Vitro: 

DMSO : 62.5 mg/mL (120.09 mM; Need ultrasonic)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 1.9215 mL 9.6073 mL 19.2145 mL
5 mM 0.3843 mL 1.9215 mL 3.8429 mL
10 mM 0.1921 mL 0.9607 mL 1.9215 mL
*Please refer to the solubility information to select the appropriate solvent.
In Vivo:
  • 1.

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% saline

    Solubility: ≥ 2.08 mg/mL (4.00 mM); Clear solution

*All of the co-solvents are provided by MCE.
References
Kinase Assay
[1]

The cells are palced in a petri dish and BCECF present in trichomonads is fixed for electron microsopy. Controns without BCECF loading, without DAB treatment, and without UV illumination are run in parallel. Trichomonads remaine viable during the procedure, except that without BCECF they are dead and disrupted by 4 h. Then the cell are lysed. After lysis, BCECF is added to the lysate to 20 μM, and the lysate is incubated,followed by 3 washes in lysis buffer to remove free dye and resuspension of the pellet in the original volume[1].

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

References

Purity: ≥96.0%

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Product Name:
BCECF
Cat. No.:
HY-101882
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