CFSE Staining Kit for PBMC staining

September 22, 2017

Anaesthesiology
LMU Munich
Researcher

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Company:

Biolegend

Product Name:

CFSE Cell Division Tracker Kit

Catalog Number:

423801

To assay long-term T-cell proliferation intensity correlating with immunoparalysis before and after surgical intervention we took advantage of this easy-to-handle assay.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMCs, Human T Cells

Primary Incubation

5µM, stock diluted in PBS, 20 Mins @ 37°C, protected from light

Blocking Agent

None

Secondary Incubation

for assessment of T-cell proliferation: 5 days, assay-dependent

Tertiary Incubation

None

Detection

Flow Cytometry

Results Summary

The cells were fluorescently labeled at day 1and stimulated with IL2 and anti CD3/CD28 beads. During proliferation, CFSE is equally distributed among the "daughter cells", whereas the overall fluorescence in each proliferating cell decreases. Therefore, CFSE staining can be used as a quick screening tool for cellular proliferation of virtually all cell entities.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Affordable, fast incubation period, easy-to-use method, cheap

The Bad

Be sure to protect the cells from light during incubation (photobleaching), CFSE ist cytotoxic to some cells!

The Bottom Line

Quick protocol, easy to handle, possibility of higher throughput than other assays (BrdU incorporation), good screening method

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