APC Anti-CD115 Antibody

March 06, 2018

Pediatrics
UW-Madison
Postdoc

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Quality of Results

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Company:

BioLegend

Product Name:

APC anti-mouse CD115 (M-CSFR)

Catalog Number:

135509

I use anti-CD115 (M-CSF receptor) to aid in labeling monocyte lineage cells. I titrated the APC-conjugated antibody on bone marrow leukocytes to determine the best concentration to test. I followed up by staining bone marrow cells that had been differentiated with GM-CSF and IL-4 for 6-7 days.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse bone marrow leukocytes (either before or after differentiation culture)

Primary Incubation

Dilutions of antibody as indicated in staining cocktail. Incubated at 4C for 30 minutes.

Blocking Agent

Medium contained FcBlock and 0.5% BSA

Secondary Incubation

Fixed cells with 2% paraformaldehyde after washing.

Tertiary Incubation

none

Detection

Fluorescence

Results Summary

For bone marrow leukocytes I gated monocytes as CD11b+, Ly6C+, Ly6G- and looked at CD115 staining. Only a small proportion of the monocytes expressed CD115. Only a small improvement in staining occurred when increasing the concentration of antibody. It appeared that the concentration with the highest amount of CD115+ cells without too much background was 1:50. I used APC anti-CD115 again in a cocktail staining bone marrow leukocytes differentiated for 7 days with GM-CSF and IL-4 (for BMDCs). Shown are CD11b+ Ly6G- cells from a single experiment. The experiment shown demonstrates an expected result where a large proportion are visibly CD115+. This result has been variable from experiment to experiment where sometimes there is no visible difference between positive and negative populations.

DOI or PMID #

N/A

Additional Notes

Anti-CD115 doesn't stain cells very brightly, this should be concedered when selecting a fluorescent conjugate and when setting flow cytometer voltages.

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Summary

The Good

Works to stain for CD115 (M-CSFR)

The Bad

Some variablity in the clarity of results.

The Bottom Line

CD115 by itself isn't a great marker to mark monocyte lineage cells but works well in tandem with other surface markers like Ly6C and CD11b.

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