Antibody of Choice for Anti-Mouse F4/80

October 09, 2013

Overall

Quality of Results

Ease-of-Optimization

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CVD
Mayo Clinic
Research Fellow

Company:

Biolegend

Product Name:

Anti-mouse F4/80 BV421

Catalog Number:

123132

I used this antibody to identify macrophages from Aorta and Kidney. For Aorta, I titrated the concentration to 3ul for 10^6 cells in 100ul and for Kidney I use around 5ul for 10^6 cells in 100ul. The reason for using this antibody is, BV421 is a bright fluorophore and cell populations are distinctly visible. I have used F4/80 tagged to FITC and Percp-cy5.5. Although, F4/80 Percp-cy5.5 is great antibody, the Percp-cy5.5 fluorophore has a wide emission range and has a potential of creating problems in multi-color flow experiments. BV421 compensates well with other BV711 and eF605NC and eF650NC. The compensation is tricky with UV live-dead stain (Life Tech) since both BV421 and the UV live-dead use 450/50 filter. However, this is a known issue.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse kidney and aorta

Primary Incubation

45 min

Blocking Agent

FCR block

Secondary Incubation

none

Tertiary Incubation

none

Detection

flow cytometry (407 laser; 450/50 filter)

Results Summary

F4/80+ macrophages stain bright and can be seen as distinct population both in aorta and kidney. They co-localize with CD11b.

Additional Notes

Make sure you titrate the antibody concentration.

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Summary

The Good

Excellent antibody for mouse F4/80+ macrophages

The Bad

Costly; BV21 does not compensate easily with UV live-dead dyes since both (UV and BV421) use 450/50 filter.

The Bottom Line

I use this regularly for multi-color experiments. Its a reliable product.

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