CFSE

Catalog No.S8269 Batch:S826901

Technical Data

Formula

C₂₉H₁₉NO₁₁

Molecular Weight

557.46

CAS No.

150347-59-4

Solubility (25°C)*

In vitro

DMSO

100 mg/mL (179.38 mM)

Water

Insoluble

Ethanol

Insoluble

In vivo (Add solvents to the product individually and in order)

Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	2.5mg/ml	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	5.0mg/ml	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.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description	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.
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.
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].

Protocol (from reference)

Cell Assay:^{^[1]}	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.
Animal Study:^{^[2]}	Animal Models C57Bl/6 mice Dosages 10 μM Administration injected into thymic lobe

Cell Assay:^{^[1]}

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.

Animal Study:^{^[2]}

Animal Models
C57Bl/6 mice
Dosages
10 μM
Administration
injected into thymic lobe

References

Selleck's CFSE has been cited by 19 publications

Local TSH/TSHR signaling promotes CD8+ T cell exhaustion and immune evasion in colorectal carcinoma [ Cancer Commun (Lond), 2024, 10.1002/cac2.12605]	PubMed: 39285586
Platelet extracellular vesicles preserve lymphatic endothelial cell integrity and enhance lymphatic vessel function [ Commun Biol, 2024, 7(1):975]	PubMed: 39128945
NLRC5 potentiates anti-tumor CD8+ T cells responses by activating interferon-β in endometrial cancer [ Transl Oncol, 2023, 36:101742]	PubMed: 37531863
NLRC5 potentiates anti-tumor CD8+ T cells responses by activating interferon-β in endometrial cancer [ Transl Oncol, 2023, 10.1016/j.tranon.2023.101742]	PubMed: 37531863
A Temporal PROTAC Cocktail-Mediated Sequential Degradation of AURKA Abrogates Acute Myeloid Leukemia Stem Cells [ Adv Sci (Weinh), 2022, e2104823]	PubMed: 35652200
ATF3 Positively Regulates Antibacterial Immunity by Modulating Macrophage Killing and Migration Functions [ Front Immunol, 2022, 13:839502]	PubMed: 35370996
Viperin deficiency promotes dendritic cell activation and function via NF-kappaB activation during Mycobacterium tuberculosis infection [ Inflamm Res, 2022, 10.1007/s00011-022-01638-3]	PubMed: 36315280
Venetoclax enhances NK cell killing sensitivity of AML cells through the NKG2D/NKG2DL activation pathway [ Int Immunopharmacol, 2022, 104:108497]	PubMed: 34999394
Chrysin Enhances Anti-tumor Immunity Response through IL-12-STAT4 Signal Pathway in B16F10 Melanoma Mouse Model [ Scand J Immunol, 2022, e13177]	PubMed: 35484925
Blockade of AIM2 inflammasome or α1-AR ameliorates IL-1β release and macrophage-mediated immunosuppression induced by CAR-T treatment [ J Immunother Cancer, 2021, 9(1)e001466]	PubMed: 33414262

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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