Labeling CCR2 on Monocytes

January 14, 2016

Anaesthesiology
LMU Munich
Researcher

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Company:

BioLegend

Product Name:

PE anti-human CD192 (CCR2) Antibody

Catalog Number:

357205

We used this antibody in order to label CCR2-expression on human monocytes while studying migrative and invasive behavior in response to stimuli such as LPS and Interferon gamma.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human Monocytes

Primary Incubation

2 million in 100µl, 2µl ab/test, 20 Mins on ice in the dark, wash 3 times

Blocking Agent

Tru Stain FcX (BioLegend), 10 Mins room temperature (5µl/100µl)

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow Cytometry

Results Summary

After incubation of monocytes in FACS-Buffer (PBS, 1%BDS, 0,01% NaAzide) for 20 Mins on ice in the dark, reliable staining of CCR2 on human monocytes cultivated for 24h/freshly isolated monocytes was obtained.

Additional Notes

Be sure to Fc-Block samples (10 Minutes, room temperature)

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Summary

The Good

Quick protocol, clear directions, antibody dilution as described works properly

The Bad

3 times washing might be a little too much for sensitive cells

The Bottom Line

Quick, reliable staining, reproducible, easy to use, cost-effective, clear discrimination of CCR2 positive and negative cells,

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