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Reagentes e instrumentos para imunologia, biologia celular e biologia molecular.

CFDA-SE [150347-59-4]

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Referência HY-D0938-5mg

Tamanho : 5mg

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Marca : MedChemExpress

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Telefone : +1 850 650 7790

CFDA-SE is a fluorescent dye that can penetrate the cell membrane. It can react with the free amine group in the cytoskeleton protein inside the cell, and finally form a protein complex with fluorescence. After entering the cell, CFDA-SE locates in the cell membrane, cytoplasm and nucleus, and the fluorescence staining is strongest in the nucleus. CFDA-SE dye can be uniformly inherited by the cells with cell division and proliferation, and its attenuation is proportional to the number of cell divisions. This phenomenon can be detected and analyzed by flow cytometry under the excitation light of 488 nm, and can be used to detect the proliferation of cells.

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CFDA-SE Chemical Structure

CAS No. : 150347-59-4

This product is a controlled substance and not for sale in your territory.

Based on 70 publication(s) in Google Scholar

: CFDA-SE purchased from MedChemExpress. Usage Cited in: Proc Natl Acad Sci U S A. 2022 Aug 9;119(32):e2201899119. [Abstract]; Flow cytometry analysis of CD4+ T cell number (B), proliferation rate (C), and representative images of CFSE-based proliferation assay (D) of CD4+ T cells cocultured with AT2 cells from healthy, LLC-bearing GPX3fl/fl, or GPX3cKO mice respectively.

: CFDA-SE purchased from MedChemExpress. Usage Cited in: Cancer Lett. 2022 May 17;541:215746. [Abstract]; PBMCs are labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) (2.5 μM). The proliferation of T cells is assessed by CFSE intensity using flow cytometry.

: CFDA-SE purchased from MedChemExpress. Usage Cited in: OncoImmunology. 2022 Feb 1;11(1):2032918. [Abstract]; Bone marrow cells are incubated with 5 μΜ CFSE (5, 6- carboxyfluorescein diacetate) for 10 min at 37°C.

: CFDA-SE purchased from MedChemExpress. Usage Cited in: Theranostics. 2021 Feb 20;11(9):4251-4261. [Abstract]; Mouse MC38 cells are washed with PBS and labeled with 1 μM CFDA-SE in PBS for 30 minutes at room temperature.

: CFDA-SE purchased from MedChemExpress. Usage Cited in: J Cell Mol Med. 2021 Jul;25(14):7052-7065. [Abstract]; THP-1 cells are stained with 1 μM carbox fluorescenceindiacetate succinimidyl ester (CFSE) in 0.1% BSA for 10 minutes at 37°C and ended with PBS for 10 minutes.

: CFDA-SE purchased from MedChemExpress. Usage Cited in: Mol Nutr Food Res. 2022 Jan;66(2):e2100619. [Abstract]; CFDA-SE (10 μM) is added to bacterial suspension and then incubated at 37°C in the dark for 20 min. The four strains are labeled with fluorescent probe CFDA-SE and were gavaged to baby rats to evaluate their colonization ability. Compared to the control group, the CFDA-SE fluorescence is present in rats gavaged with the four strains.

Description

CFDA-SE is a fluorescent dye that can penetrate the cell membrane. It can react with the free amine group in the cytoskeleton protein inside the cell, and finally form a protein complex with fluorescence. After entering the cell, CFDA-SE locates in the cell membrane, cytoplasm and nucleus, and the fluorescence staining is strongest in the nucleus^[1]. CFDA-SE dye can be uniformly inherited by the cells with cell division and proliferation, and its attenuation is proportional to the number of cell divisions. This phenomenon can be detected and analyzed by flow cytometry under the excitation light of 488 nm, and can be used to detect the proliferation of cells^[1].

In Vitro

Preparation of CFDA-SE working solution
1.1 Preparation of the stock solution
Dissolve 1 mg of CFDA-SE in 0.1794 mL of DMSO to obtain 10 mM of CFSE.
Note: It is recommended to store the stock solution at -20 ℃ or -80 ℃ away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of CFDA-SE working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 5-10 μM of CFDA-SE working solution.
Note: Please adjust the concentration of CFDA-SE working solution according to the actual situation.
Cell staining
2.1 For suspension cells: Centrifuge at 1000 g at 4℃ 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℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of CFDA-SE working solution, and then incubate at room temperature for 30 minutes.
2.3 Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

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

Masse moléculaire

557.46

Formule

C₅₈H₃₈N₂O₂₂

CAS No.

150347-59-4

Appearance

Solid

Color

Off-white to yellow

Emission (Em)

515

Excitation (Ex)

485

SMILES

CC(OC1=CC=C(C2(C(C=CC(C(ON3C(CCC3=O)=O)=O)=C4)=C4C(O2)=O)C(C=CC(OC(C)=O)=C5)=C5O6)C6=C1)=O.CC(OC7=CC=C(C8(C(C=C(C(ON9C(CCC9=O)=O)=O)C=C%10)=C%10C(O8)=O)C(C=CC(OC(C)=O)=C%11)=C%11O%12)C%12=C7)=O

Livraison

Room temperature in continental US; may vary elsewhere.

Stockage

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvant et solubilité

In Vitro:

DMSO : 50 mg/mL (89.69 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	1.7939 mL	8.9693 mL	17.9385 mL
5 mM	0.3588 mL	1.7939 mL	3.5877 mL
10 mM	0.1794 mL	0.8969 mL	1.7939 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 (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Calculateur de molarité
Calculateur de dilution

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: ≥ 2.08 mg/mL (3.73 mM); Clear solution

This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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% Corn Oil
Solubility: ≥ 2.08 mg/mL (3.73 mM); Clear solution

This protocol yields a clear solution of ≥ 2.08 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 (20.8 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 (sealed storage, away from moisture and 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.

Pureté et documentation

Purity: 99.01%

Select Batch:

SDS (393 KB)

COA (251 KB) HNMR (330 KB) LCMS (264 KB)

Instruction de manipulation (2659 KB)

Dyeing Example

Description: CFDA-SE is an intracellular fluorescent dye with green fluorescence, it can be used to detect strain colonization in vivo.
Method: For determine strain colonization in the intestine.
1. Strains are re-suspended in PBS to obtain a cell density of 10⁸ CFU/mL.
2. CFDA-SE (1 mM; 10 μL; 20 min; 37°C; dark) is added to 1 mL of bacterial suspension.
3. Centrifuge (13000 g, 10 min, 4°C) and wash mixture for three times using sterile PBS to remove excess CFDA-SE.
4. Strains are then re-suspended in sterile PBS (10⁸ CFU/mL) and 1 mL of bacterial suspension is gavaged to SD rats. Day 1 and 3 after gavage, the fluorescence imaging is performed on anesthetized rats who are then sacrificed and different intestinal sections are taken for imaging.
5. The fluorescence imaging is performed using an IVIS Lumina III Smart Imaging System (PerkinElmer, USA).
Description: CFDA-SE is an intracellular fluorescent dye with green fluorescence, it can be used to stain bacteria.
Method: For bacteria adhesion assay.
1. Testing cells are inoculated into confocal laser dishes at a density of 1×10⁴ cells per well and cultured for 24 h.
2. The bacteria suspension is resuspended with CFDA-SE, then centrifuged with aseptic PBS.
3. Cultured cells are infected with CFDA-SE labeled bacteria in the presence of drug solutions in dark for 2 h.
4. Finally, the dishes are placed under confocal laser scanning microscopy (FV3000, Japan) for inspection.
Description: CFDA-SE is an intracellular fluorescent dye with green fluorescence, it can be used to assess cell binding.
Method: For monocyte adhesion assay.
1. Cells are labeled with CFDA-SE (5 μM; 4 h; 37°C) with the HUVECs in a moist atmosphere containing 5% CO₂.
2. Unbound cells in the dishes are removed by washing 5 times with serum-free medium.
3. Adherent cells are visualized under an inverted fluorescence microscope (Olympus IX71, Japan).
Description: CFDA-SE is an intracellular fluorescent dye with green fluorescence, it can be used to detect cell membrane fusion.
Method: For monocyte adhesion assay.
1. Testing cells are stained with two different dyes (including CFDA-SE), separately.
2. Fusion is induced by adding drops of pre-warmed 50% PEG 2000 at 37°C. PEG fusion is terminated by adding pre-warmed DMEM medium containing 15% serum to the tube after one minute. After being centrifuged at 1000 g for 5 min, testing fused cells are obtained.
3. Use a fluorescence microscope for image.
Description: CFDA-SE is an intracellular fluorescent dye with green fluorescence, it can be used to determine cell lethality.
Method: For cell staining (Flow cytometric detection of γδT cell killing rate).
1. CFDA-SE (10 mg) is precisely added to 1 mL of DMSO solution to prepare a solution with a concentration of 17.9385 mM and diluted according to actual needs.
2. Culture cells with CFDA-SE at a ratio of 10:1.
3. Wash cells with PBS and use a microscope for image.

Références

[1]. Banks HT, et al. Quantifying CFSE Label Decay in Flow Cytometry Data. Appl Math Lett. 2013 May 1;26(5):571-577. [Content Brief]

[2]. A Bruce Lyons, et al. Flow cytometric analysis of cell division by dilution of CFSE and related dyes. Curr Protoc Cytom. 2013;Chapter 9:Unit9.11. [Content Brief]

[3]. Cui J, et al. Prostaglandin E3 attenuates macrophage-associated inflammation and prostate tumour growth by modulating polarization. J Cell Mol Med. 2021 Jun;25(12):5586-5601. [Content Brief]

[4]. Xiuyun Jiang et al. Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment. Oncoimmunology. 2017; 6(7): e1333210. [Content Brief]

Test cellulaire ^[2]	Single-cell suspensions of splenocytes (or PBMC) are stained with 1 µM CFSE in PBS for 9 min at 37°C, combined with 20 mL of 10% FBS RPMI-1640 medium at RT for 2 min, centrifuged, washed, and counted. 2×10⁶ CFSE-labeling splenocytes are added to the top of the slices in 24-well membrane culture insert^[2]. MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.
Références	[1]. Banks HT, et al. Quantifying CFSE Label Decay in Flow Cytometry Data. Appl Math Lett. 2013 May 1;26(5):571-577. [Content Brief] [2]. A Bruce Lyons, et al. Flow cytometric analysis of cell division by dilution of CFSE and related dyes. Curr Protoc Cytom. 2013;Chapter 9:Unit9.11. [Content Brief] [3]. Cui J, et al. Prostaglandin E3 attenuates macrophage-associated inflammation and prostate tumour growth by modulating polarization. J Cell Mol Med. 2021 Jun;25(12):5586-5601. [Content Brief] [4]. Xiuyun Jiang et al. Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment. Oncoimmunology. 2017; 6(7): e1333210. [Content Brief]

Test cellulaire
^[2]

Single-cell suspensions of splenocytes (or PBMC) are stained with 1 µM CFSE in PBS for 9 min at 37°C, combined with 20 mL of 10% FBS RPMI-1640 medium at RT for 2 min, centrifuged, washed, and counted. 2×10⁶ CFSE-labeling splenocytes are added to the top of the slices in 24-well membrane culture insert^[2].

MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.

Références

[1]. Banks HT, et al. Quantifying CFSE Label Decay in Flow Cytometry Data. Appl Math Lett. 2013 May 1;26(5):571-577. [Content Brief]

[2]. A Bruce Lyons, et al. Flow cytometric analysis of cell division by dilution of CFSE and related dyes. Curr Protoc Cytom. 2013;Chapter 9:Unit9.11. [Content Brief]

[3]. Cui J, et al. Prostaglandin E3 attenuates macrophage-associated inflammation and prostate tumour growth by modulating polarization. J Cell Mol Med. 2021 Jun;25(12):5586-5601. [Content Brief]

[4]. Xiuyun Jiang et al. Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment. Oncoimmunology. 2017; 6(7): e1333210. [Content Brief]

Complete Stock Solution Preparation Table

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

DMSO	1 mM	1.7939 mL	8.9693 mL	17.9385 mL	44.8463 mL
	5 mM	0.3588 mL	1.7939 mL	3.5877 mL	8.9693 mL
	10 mM	0.1794 mL	0.8969 mL	1.7939 mL	4.4846 mL
	15 mM	0.1196 mL	0.5980 mL	1.1959 mL	2.9898 mL
	20 mM	0.0897 mL	0.4485 mL	0.8969 mL	2.2423 mL
	25 mM	0.0718 mL	0.3588 mL	0.7175 mL	1.7939 mL
	30 mM	0.0598 mL	0.2990 mL	0.5980 mL	1.4949 mL
	40 mM	0.0448 mL	0.2242 mL	0.4485 mL	1.1212 mL
	50 mM	0.0359 mL	0.1794 mL	0.3588 mL	0.8969 mL
	60 mM	0.0299 mL	0.1495 mL	0.2990 mL	0.7474 mL
	80 mM	0.0224 mL	0.1121 mL	0.2242 mL	0.5606 mL

CFDA-SE Related Classifications

Others

Others
Fluorescent Dye

CFDA-SE [150347-59-4]

Referência HY-D0938-5mg

Contactar o distribuidor local :

Marca : MedChemExpress

Contactar o distribuidor local :

Calculateur de molarité

Calculateur de dilution

Complete Stock Solution Preparation Table

CFDA-SE Related Classifications

Keywords:

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CFDA-SE [150347-59-4]

Referência HY-D0938-5mg

Contactar o distribuidor local :

Marca : MedChemExpress

Contactar o distribuidor local :

CFDA-SE Related Antibodies

Calculateur de molarité

Calculateur de dilution

Complete Stock Solution Preparation Table

CFDA-SE Related Classifications

Keywords:

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