Bright Enough to Separate CD4+ Cells, but Requires Compensation

August 29, 2018

Developmental Immunology
La Jolla Institute for Allergy and Immunology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

V500 Rat anti-Mouse CD4 Clone RM4-5

Catalog Number:

560782

The general focus of my work is to analyze the response different cell types in mouse lungs after pulmonary bacteria infection. A variety of fluorophore-labeled antibodies were tested for staining CD4+ T cells in the lung cells of mice.

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

The V500 flurophore is bright enough to detect CD4+ T cells from lung homogenates. However, it is notable that is is a more difficult fluorophore to compensate, and is not ideal for highly autofluorescent samples.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Clearly labels CD4+ cells, highly specific

The Bad

Not ideal for highly autofluorescent samples

The Bottom Line

Dim markers like V500 work fine for the abundant CD4 surface marker. Just not ideal for autofluorescent or homogenates with a large percentage of dead cells.

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