Overview

  • Product name
    CFSE - Cell Labeling Kit
    See all Cell labeling kits
  • Detection method
    Fluorescent
  • Assay type
    Cell-based
  • Product overview

    CFSE is a versatile tool for the fluorescent intracellular labeling of live cells. Labeled cells can be assayed using flow cytometry and fluorescent microscopy. The dye is long lasting and well retained within labeled cells. The provided CFSE is sufficient for ~1000 assays.


    CFSE is a cell permeant, non-fluorescent pro-dye. Intracellular esterases in live cells cleave the acetate groups of CFSE producing a molecule with green fluorescence that is also now membrane impermeant. The succinimidyl ester group reacts indiscriminately with intracellular free amines to generate covalent dye-protein conjugates. The result is live cells with an intracellular fluorescent label. At appropriate concentrations CFSE is non-toxic to cells and the fluorescence is retained after formaldehyde and alcohol fixation. CFSE labeled cells can be detected with any instrument or filter set compatible with FITC detection: Excitation(max)=492nm, Emission(max)=517nm.

Properties

Images

  • Duplexing treated and untreated cells in a single sample. One plate of HeLa cells was labeled with ab113853 (CFSE) and a second plate was treated with the hypoxia mimetic DFO but not labeled with CFSE. After harvesting, the cells were combined in a single tube, stained with a HIF1A antibody and subjected to flow cytometry. Expected results are shown in the left panel cartoon. The no primary antibody control sample (middle panel) shows the resolution of CFSE labeled and unlabeled cells. The HIF1A antibody stained cells (right panel) shows that only cells exposed to DFO have an increase in HIF1A protein levels.

  • Flow cytometry analysis of ab113853 (CFSE) dilution with cell division. Jurkat cells were labeled with 1µM CFSE on day0 and then a portion of the culture was subjected to flow cytometry analysis on days 1-7. Shown, an overlay histogram of the daily CFSE fluorescence intensity.
  • Jurkat cells labeled with ab113853 (CFSE). Jurkat cells were labeled with 1mM CFSE in media for 15 minutes, washed once with PBS and imaged on a flurorescence microscope. The cells in this image are live but fixed cells give similar results.

Protocols

References

This product has been referenced in:
  • Li Z  et al. Reiterative Enrichment and Authentication of CRISPRi Targets (REACT) identifies the proteasome as a key contributor to HIV-1 latency. PLoS Pathog 15:e1007498 (2019). Read more (PubMed: 30645648) »
  • Poh PSP  et al. Osteogenic Effect and Cell Signaling Activation of Extremely Low-Frequency Pulsed Electromagnetic Fields in Adipose-Derived Mesenchymal Stromal Cells. Stem Cells Int 2018:5402853 (2018). Read more (PubMed: 30123287) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer



I contacted the laboratories to discuss this further. They also suggest that the CFSE - Cell Labeling Kit would be easier to use than BrdU.

1. CFSE versus BrdU - these are very different assays.
BrdU is a more direct measure of proliferation – you look at DNA synthesis by BrdU incorporation. It is also a bit onerous in terms of sample prep – DNA needs to be denatured before Ab staining. BrdU assays can be read on microplates or in flow cytometer. Assay window is typically short – pulse of BrdU and looking at DNA synthesis over a short time frame.

CFSE is a less direct but very simple assay – label cells, wash away dye, examine dye dilution over time (as cells divide the dye is diluted – less dye per cell = dividing cells). These are longer timepoint assays, typically days post labeling. Needs flow cytometer for analysis.



2. How much to use?

Anywhere from 10nM – 10uM. Needs to be determined by the user. Amounts used in the sample data are given in the booklet – these could be the starting concentrations to test.

From the product booklet page 9-10.

Concentrations of CFSE staining will need to be determined individually based on the cell line used

and the goals of the experiment. For example, if following cell proliferation by CFSE dilution, higher

levels of CFSE staining is desired. If using CFSE to label cells as a counterstain for additional

fluorescent markers, lower levels of CFSE staining would prevent fluorescent spillover between filter

sets.

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Answer

Thank you for contacting us.

No stimulation was performed in this assay, simply culturing the cells in standard growth media.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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