6-CFDA (Synonyms: 6-Carboxyfluorescein diacetate)

Cat. No.: HY-D0721 Purity: 98.09%: FAQs
Data Sheet SDS COA Handling Instructions

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6-CFDA is a common aliphatic luciferin-line organism. CFDA conducts free diffusion into cells, and then it is hydrolyzed into carboxyl fluorescein (CF) by intracellular non-specific lipase. CF containing portion contains an additional negative charge so that it is better retained in cells, compared to fluorescein dyes.

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

6-CFDA Chemical Structure

CAS No. : 3348-03-6

Size	Price	Stock	Quantity
5 mg	USD 46	In-stock	Estimated Time of Arrival: December 31
10 mg	USD 72	In-stock	Estimated Time of Arrival: December 31
50 mg	USD 96	In-stock	Estimated Time of Arrival: December 31
100 mg	USD 168	In-stock	Estimated Time of Arrival: December 31
200 mg		Get quote
500 mg		Get quote

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Other Forms of 6-CFDA:

5-CFDA In-stock
5(6)-CFDA In-stock

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Description

In Vitro

Preparation of 6-CFDA working solution
1.Preparation of the stock solution
Dissolve 1mg 6-CFDA in 0.2172 mL DMSO to obtain 10 mM of 6-CFDA.
Note: It is recommended to store the stock solution at -20 °C -80 °C away from light and avoid repetitive freeze-thaw cycles.
2.Preparation of 6-CFDA working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of 6-CFDA working solution.
Note: Please adjust the concentration of 6-CFDA working solution according to the actual situation.

Cell staining
1.Cell preparation:
For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.Add 1 mL of 6-CFDA working solution, and then incubate at room temperature for 30 minutes.
3.Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
4.Wash twice with PBS, 5 minutes each time.
5.Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

Precautions
1. It is recommended to store the stock solution at -20℃ or -80℃ away from light and avoid repetitive freeze-thaw cycles.
2. Please adjust the concentration of 6-CFDA working solution according to the actual situation.
3. This product is for R&D use only, not for drug, household, or other uses.
4. For your safety and health, please wear a lab coat and disposable gloves to operate.

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

Molecular Weight

460.39

Formula

C₂₅H₁₆O₉

CAS No.

3348-03-6

Appearance

Solid

Color

White to off-white

Emission (Em)

517

Excitation (Ex)

492

SMILES

CC(OC1=CC=C(C2(O3)C4=CC(C(O)=O)=CC=C4C3=O)C(OC5=C2C=CC(OC(C)=O)=C5)=C1)=O

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 : 100 mg/mL (217.21 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	2.1721 mL	10.8604 mL	21.7207 mL
5 mM	0.4344 mL	2.1721 mL	4.3441 mL
10 mM	0.2172 mL	1.0860 mL	2.1721 mL

View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (5.43 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (5.43 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Protocol 3

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (5.43 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

DMSO +

Tween-80 +

Saline

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Calculation results:

Working solution concentration: mg/mL

Method for preparing stock solution: mg drug dissolved in μL DMSO (Stock solution concentration: mg/mL).

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

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).

Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.

If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.

Purity & Documentation

Purity: 98.09%

Select Batch:

Data Sheet (278 KB) SDS (393 KB)

COA (254 KB) LCMS (249 KB)

Handling Instructions (2659 KB)

References

[1]. Yang T, et al. A novel nonradioactive CFDA assay to monitor the cellular immune response in myeloid leukemia. Immunobiology. 2013 Apr;218(4):548-53. [Content Brief]

[2]. Card SD, et al. Assessment of fluorescein-based fluorescent dyes for tracing Neotyphodium endophytes in planta. Mycologia. 2013 Jan-Feb;105(1):221-9. [Content Brief]

[3]. Fang X, et al. Bone marrow-derived endothelial progenitor cells are involved in aneurysm repair in rabbits. J Clin Neurosci. 2012 Sep;19(9):1283-6. [Content Brief]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

DMSO	1 mM	2.1721 mL	10.8604 mL	21.7207 mL	54.3018 mL
	5 mM	0.4344 mL	2.1721 mL	4.3441 mL	10.8604 mL
	10 mM	0.2172 mL	1.0860 mL	2.1721 mL	5.4302 mL
	15 mM	0.1448 mL	0.7240 mL	1.4480 mL	3.6201 mL
	20 mM	0.1086 mL	0.5430 mL	1.0860 mL	2.7151 mL
	25 mM	0.0869 mL	0.4344 mL	0.8688 mL	2.1721 mL
	30 mM	0.0724 mL	0.3620 mL	0.7240 mL	1.8101 mL
	40 mM	0.0543 mL	0.2715 mL	0.5430 mL	1.3575 mL
	50 mM	0.0434 mL	0.2172 mL	0.4344 mL	1.0860 mL
	60 mM	0.0362 mL	0.1810 mL	0.3620 mL	0.9050 mL
	80 mM	0.0272 mL	0.1358 mL	0.2715 mL	0.6788 mL
	100 mM	0.0217 mL	0.1086 mL	0.2172 mL	0.5430 mL

6-CFDA Related Classifications

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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:

6-CFDA3348-03-66-Carboxyfluorescein diacetateFluorescent DyeflowcytometryfluorescencemicroscopystainInhibitorinhibitorinhibit

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