CCR6-Labeling on Human Primary PBMCs

July 05, 2016

Anaesthesiology
LMU Munich
Researcher

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Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

FITC anti-human CD196 (CCR6) Antibody

Catalog Number:

353411

We included this antibody in a FACS panel discriminating T-cells. In this panel, we looked for Th1 Th2 and Treg populations and their differences in human primary PBMC.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Primary Human PBMC

Primary Incubation

5µl per 1 million cells, incubation 20 Mins on ice in the dark, wash 3 times with FACS buffer (PBS, 1% BSA, 0,001% Na Azide)

Blocking Agent

Tru Stain FcX Receptor blocking solution (Biolegend)

Secondary Incubation

None

Tertiary Incubation

None

Detection

Flow Cytometry, Attune Acoustic Focusing Cytometer

Results Summary

This antibody may be used for labeling CCR6-positive cells. Although the populations are a little blurry, this antibody gives reproducible results and is relatively resistant to photobleaching.

Additional Notes

Be sure to Fc Block prior toadding the antibody. When using ledd than 1 million cells you may also reduce the amount of antibody.

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Summary

The Good

Easy to use, recommended titration works. Inexpensive, rather resistant to photobleaching

The Bad

Repeated washing times.

The Bottom Line

Robust and easy to use, good antibody, even useable for rather weak expressions of CCR6 (e.g. on human PBMCs), results are reproducible, recommended antibody concentration works well on PBMC.

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