Anti-murine F4/80 FITC

May 01, 2017

Pediatrics
UW-Madison
Postdoc

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Biolegend

Product Name:

FITC anti-mouse F4/80 Antibody

Catalog Number:

123107

F4/80 is a marker found on macrophages. For purposes of staining macrophages in multiple tissues, I titrated anti-F4/80-FITC on splenocytes to stain for splenic macrophages (from naive mouse). I included other markers in the stain cocktail in this experiment to aid in identification and visualization of the data.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Splenocytes from naive C57B/6 mice

Primary Incubation

Titrations (as shown) for 30 minutes at 4C along with antibodies against other markers for gating.

Blocking Agent

Buffer contained BSA and FcBlock

Secondary Incubation

None

Tertiary Incubation

None

Detection

Fluoresecence in flow cytometry

Results Summary

The population of splenocytes I interrogated for anti-F4/80 expression were B220-, CD11c- splenocytes. The F4/80+ splenocytes did not have very high CD11b expression in my samples (maybe due to the fact that the spleens collected were from naive mice). The F4/80 marker alone does not appear to be enough to distinguish the population of macrophages (at least for this product). The population of F4/80+ cells was small in naive spleens, but was distinct enough to identify with minimal pre-gating.Notably, the F4/80+ population shown was distinct from the F4/80- population even when staining cells with a 1:400 dilution of the antibody.

DOI or PMID #

N/A

Additional Notes

Macrophages tend to be more autofluorescent (mostly in the FITC channel) than most other cells, especially when activated. Depending on the application of this antibody, using the FITC conjugate may be beneficial or detrimental. If the design is to gate macrophages generally, this particular reagent (FITC conjugate) will be benfitted by the fact that the cell they stain will have extra fluorescence in the FITC channel. This would be a great reagent to gate out macrophages, but would not be good if you wanted to tease out macrophage populations or determine relative F4/80 expression on macrophages.

Related Categories

Image Gallery

Summary

The Good

Stains consistently between 1:100 and 1:400 dilutions. F4/80+ population distinguishable from F4/80- population.

The Bad

Anti-F4/80 alone is probably not enough in an experiment investigating macrophages.

The Bottom Line

This product is a good one if you desire to stain for F4/80 in your experiment. The use of the FITC (or similiar Alexa488) conjugate may be beneficial, but could hinder your analysis.

Share your experience with other scientists. Write a Review! »

Join the discussion