Size | Price | Stock | Qty |
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1mg |
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5mg |
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10mg |
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25mg |
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Other Sizes |
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BCECF is a novel fluorescent agent/dye which a pH-sensitive and has been widely used as an indicator of cytosolic pH. It can be used for measurements in the physiological pH range 6.0–8.0. It has a λexcitation of 420~520 nm and λemission of 530 nm.
ln Vitro |
Advice (This is our suggested protocol, which should be adjusted to suit your particular circumstances as it simply offers guidance). 1. Prepare a stock solution of 2–20 mM BCECF in DMSO and store it at -20°C. 2. Make a buffer solution (5–50 μM BCECF dye loading) using Hepes (pH 7.4) or PBS. 3. To put the BCECF dye in the cell plate, add 1000 μL/well (6-well plate), 100 μL/well (96-well plate), or 25 μL/well (384-well plate). 4. The dye-loaded plate should be in the cell culture incubator for around half an hour. 5. Rinse and use buffer in place of dye loading solution. 6. For ratiometric measurements, watch fluorescence at Ex/Em = 490/535 nm or 430/535 nm to perform a pH determination.
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References |
[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.
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Molecular Formula |
C27H20O11
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Molecular Weight |
520.4411
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CAS # |
85138-49-4
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SMILES |
O=C(O)CCC1=CC(C23OC(C4=C2C=CC(C(O)=O)=C4)=O)=C(OC5=C3C=C(CCC(O)=O)C(O)=C5)C=C1O.O=C(O)CCC6=CC(C78OC(C9=C7C=C(C(O)=O)C=C9)=O)=C(OC%10=C8C=C(CCC(O)=O)C(O)=C%10)C=C6O
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Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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Solubility (In Vitro) |
DMSO : ~62.5 mg/mL (~120.09 mM)
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Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.00 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.) |
Preparing Stock Solutions | 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 |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration: mg/mL;
Method for preparing DMSO stock solution: mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation::Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.