Staining BAL With CCR2: Easy Differentiation Of Epithelial And Monocyte Subpopulations

October 17, 2016

Medicine
University of Colorado Denver
Professional Research Assistant

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

PE anti-mouse CD192 (CCR2) from Biolegend

Catalog Number:

150609

We study innate immunology in the lung in murine models. We wanted to be able to differentiate between subpopulations of various monocytes, macrophages, and epithelial cells in the lungs via flow cytometry, and the addition of a CCR2 antibody to our panel has helped enormously with this. The BioLegend PE anti-mouse CD192 (CCR2) antibody stains well, requires little optimization, and has given us clear results for different CCR2 high and low subpopulations in the lung. We chose this product initially because it was one of the best/cheapest CCR2 antibodies, and were very pleased with the both the quality and price of the product as well as the customer service we've gotten from BioLegend.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine bronchoalveolar lavage

Primary Incubation

Cells are collected, spun, and counted, then incubated for 30mins on ice or at 4C in antibody master mix. Antibodies are diluted 1:200 or 1:400 (as per packaging on antibody) in 1X PAB buffer (BSA, NaN3 and PBS)

Blocking Agent

Cells washed 2x with 1x PAB (BSA, NaN3, PBS)

Secondary Incubation

FACS Fixative, 30 mins on ice/at 4C

Tertiary Incubation

No tertiary incubation

Detection

Flow Cytometry

Results Summary

Flow files (FSC files). CCR2 always used in a panel with other antibodies. Gate for macrophage, monocyte, and epithelial cell populations. Use CCR2 to help distinguish CCR2 + hi and lo and CCR2 - populations.See data below for an example:

Additional Notes

None

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Summary

The Good

It is reasonably priced, stains well, and required little optimization for our purposes (simple diluting according to the concentration on the tube worked fine)

The Bad

It only comes in two colors: PE and PE-Cy7. It is not always easy to work into a complicated panel.

The Bottom Line

Great product for our purposes.

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