Specific Staining for Neutrophils

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:

Alexa Fluor® 700 anti-mouse Ly-6G Antibody (Clone 1A8)

Catalog Number:

127622

The general focus of my work is to analyze the response different cell types in mouse lungs after pulmonary bacteria infection. This antibody was used to specifically stain neutrophils from mouse lungs for flow cytometry.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Collagenase-digested mouse lung tissue, including neutrophils

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

AF700 is a dim marker, but due to the high expression of surface Ly6G this antibody is adequate for good separation of Ly6G+ neutrophils.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Bright staining, specific for neutrophils

The Bad

None

The Bottom Line

Clear, specific staining. Good choice for the relatively dim AF700 fluorophore.

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