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CFSE

Name: CFSE
Brand: Selleck Chemicals
SKU: S8269
Rating: 5 (17 reviews)

Synonyms: Carboxyfluorescein succinimidyl ester, 5(6)-Carboxyfluorescein diacetate succinimidyl ester, CFDA-SE, 5(6)-CFDA N-succinmidyl ester

Catalog No.S8269 Purity:99.85%

Carboxyfluorescein succinimidyl ester (CFSE, 5(6)-Carboxyfluorescein diacetate succinimidyl ester, CFDA-SE, 5(6)-CFDA N-succinmidyl ester) is a fluorescent cell staining dye. CFSE is frequently used in cell proliferation assay and motility assays. CFSE is cell permeable and covalently couples, via its succinimidyl group, to intracellular molecules, notably, to intracellular lysine residues and other amine sources.

CFSE Chemical Structure

CAS No. 150347-59-4

Purity & Quality Control

Batch: Purity: 99.85%

99.85

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Biological Activity

Description

In vitro
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.
Cell Research	Cell lines	human erythroleukaemic cell line K562, mouse lymphoma cell line YAC-1, human mammary cancer cell line MCF-7 and human melanoma cell line A375
	Concentrations	2 µM, 3 µM, 4 µM, 5 µM, 10 µM and 20 µM
	Incubation Time	5, 6, 7, 8, 10 and 15 min
	Method	The 5 mM CFDA-SE stock in DMSO is diluted to different concentrations(2 µM, 3 µM, 4 µM, 5 µM, 10 µM and 20 µM) in PBS with a total volume of 1 ml. After each cell line is harvested and washed three times with PBS, 1×10⁹ cells are added to equal volume of CFDA-SE with different concentrations and incubated at 37°C for 5, 6, 7, 8, 10 and 15 min with agitation. The labeling reaction is stopped for 1 min by adding an equal volume of heat inactivated fetal bovine serum. The CFDA-SE labeled cells are washed twice with PBS and recounted, and the cell concentration is adjusted to 6×104cells/ml in IMDM containing 10% FCS.

In Vivo
In vivo	In vivo labeling of cells with CFSE is feasible with virtually no toxic effect on the labeled or adjacent cells. It has advantages in relatively long-term migration studies as demonstrated by the recovery of T-cells in the peripheral lymphoid organs^[2].
Animal Research	Animal Models	C57Bl/6 mice
	Dosages	10 μM
	Administration	injected into thymic lobe

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
Clinical Trial Information (data from https://clinicaltrials.gov, updated on 2024-05-22)
NCT02028403	Completed	HIV Infections	National Institute of Allergy and Infectious Diseases (NIAID)	June 2014	Phase 1
NCT01376726	Completed	HIV Infections	National Institute of Allergy and Infectious Diseases (NIAID)\|Novartis Vaccines and Diagnostics Inc	July 2011	Phase 1
NCT01029548	Completed	HIV Infections	Barbara Ensoli MD\|Istituto Superiore di Sanità	April 2008	--
NCT01024556	Completed	HIV Infection	Barbara Ensoli MD\|Istituto Superiore di Sanità	March 2008	--

References

Chemical Information & Solubility

Molecular Weight	557.46	Formula	C₂₉H₁₉NO₁₁
CAS No.	150347-59-4	SDF	Download CFSE SDF
Smiles	CC(=O)OC1=CC=C2C(=C1)OC3=C(C=CC(=C3)OC(C)=O)C24OC(=O)C5=CC(=CC=C45)C(=O)ON6C(=O)CCC6=O.CC(=O)OC7=CC=C8C(=C7)OC9=C(C=CC(=C9)OC(C)=O)C8%10OC(=O)C%11=CC=C(C=C%10%11)C(=O)ON%12C(=O)CCC%12=O
Storage (From the date of receipt)	3 years -20°C(in the dark) powder	Storage of Stock Solutions Aliquot the stock solutions to avoid repeated freeze-thaw cycles	1 year-80°Cin solvent
Storage (From the date of receipt)	3 years -20°C(in the dark) powder		1 month-20°Cin solvent

In vitro
Batch:

DMSO : 100 mg/mL ( (179.38 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molecular Weight Calculator

In vivo Batch: Add solvents to the product individually and in order.					In vivo Formulation Calculator
	Clear solution	5% DMSO 1 95% Corn oil	5.0mg/ml (8.97mM)	Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.	In vivo Formulation Calculator
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	2.5mg/ml (4.48mM)	Taking the 1 mL working solution as an example, add 50 μL of 50 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	2.5mg/ml (4.48mM)	Taking the 1 mL working solution as an example, add 50 μL of 50 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil	0.5mg/ml (0.90mM)	Taking the 1 mL working solution as an example, add 50 μL of 10 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.

Preparing Stock Solutions

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
=	×	×

Dilution Calculator Molecular Weight Calculator

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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