Clear ICOS Labeling, But Shows a Gradient

Developmental Immunology
La Jolla Institute for Allergy and Immunology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

PE/Cy7 anti-human/mouse/rat CD278 (ICOS) Antibody

Catalog Number:

313520

Our goal analyze the response different cell types in mouse lungs after pulmonary bacteria infection. Mouse lungs were digested with collagenase, strained, and stained for flow cytometry with a cocktail of different antibodies. The samples were analyzed on a BD LSRII flow cytometry. The FACS plot shown are after gating on Single, Live, CD45+, CD19+ Cells

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Collagenase-digested mouse lung tissue

Primary Incubation

1:200 for 20 minutes on ice

Blocking Agent

24G2 in 3% BSA in PBS for 10 minutes on ice prior to staining in 3% BSA

Secondary Incubation

None

Tertiary Incubation

None

Detection

Flow cytometer (BD LSRII)

Results Summary

This antibody stained specifically, and a + vs - population was detectable. However, it is notable that this staining appears as more of a gradient than a isolated population. A good FMO or isotype control is useful for this antibody.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Specific and able to separate + and - populations

The Bad

Gradiant, but more due to marker than the antibody

The Bottom Line

Would recommend this antibody and fluorophore choice for ICOS

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