Good Antibody for Staining CD4+ T Cells

August 28, 2018

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 CD4 Antibody

Catalog Number:

100430

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 label CD4+ T cells in mouse lungs. Multiple different fluorophores were tested for this application.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Collagenase-digested mouse lung tissue, including T cells

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 CD4+ cells specifically, with very clear separation of cells. AF700 is a relatively dim fluorophore, but due to the high abundance of surface CD4 good separation was achievable.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Clear and specific staining, narrow histogram peaks

The Bad

Not the brightest fluorophore, but sufficient for the application

The Bottom Line

For a large panel of antibodies, CD4 is a great choice to put on a dimmer fluorophore such as AF700.

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