Anti-IL-4 Receptor, BV421

February 04, 2019

BlueskyReddit
Pediatrics
UW-Madison
Postdoc

Overall

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Company:

BD Biosciences

Product Name:

BV421 Rat Anti-Mouse CD124

Catalog Number:

564086

Browse Similar Products: CD124 Flow Cytometry Antibodies

I titrated anti-IL4R (specific alpha subunit) to look at expression of this receptor on neutrophils. IL-4 in culture, particularly with other cytokines, has an effect on murine neutrophils so I will look to see when and to what degree the IL-4 receptor is expressed on neutrophils. This particular antibody doesn't stain well, even the representative staining plot on BD's website shows a modest increase in fluorescence over the staining control. I found similar staining with neutrophils.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine Neutrophils (derived from progenitor cells in vitro)

Primary Incubation

Antibodies at indicated amounts (other antibodies for neutrophil surface markers also included to gate neutrophils)

Blocking Agent

FcBlock (~1:75) and BSA (0.5%) were present in staining cocktail. BSA was present in wash buffers.

Secondary Incubation

Cells fixed after washing

Tertiary Incubation

N/A

Detection

Fluorescence by flow cytometry

Results Summary

Anti-IL-4R BV421 did stain the neutrophils with a surface stain. The positively staining cells were not incredibly brighter than the staining control (FMO), but with the staining control positively stained cells were seen. A 1:200 dilution of antibody (0.5 µL in 100 µL total volume) was sufficient for staining (stained ~500,000 cells).

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Can stain IL-4R

The Bad

FMO absolutely required to distinguish positive and negative populations.

The Bottom Line

This product works. Probably insufficient for use in gating populations, but with appropriate controls sufficient for investigating IL-4 receptor expression.

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