Good Marker for Sorting Naive CD4 T Cells

January 12, 2016

Research Participation Program
Oak Ridge Institute for Science and Education
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

APC Rat Anti-Mouse CD44

Catalog Number:

559250

Our goal was to sort naive CD4+ T cells from mouse spleens for a proliferation assay. We were measuring the ability of antigen presenting cells to present ovalbumin peptides to transgenic OVA-specific T cells. We selected this product in order to gate out non-naive T cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Live murine splenocytes, red blood cells lysed

Primary Incubation

1:100 dilution of each: anti-CD4 Pacific Blue, anti-CD62L Alexa Fluor 488, anti-CD44 APC. Incubate on ice 20 minutes.

Blocking Agent

1xPBS+2%FBS+2mM EDTA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

BD FACSAria II

Results Summary

T cells were gated initially using CD4. Within that population, there was a clear population of CD44low/CD62Lhigh cells that we consider to be naive T cells.

Additional Notes

None

Related Categories

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Summary

The Good

It was easy to identify the CD44low CD62Lhigh population using this color combination.

The Bad

No drawbacks observed.

The Bottom Line

This is an effective way of identifying naive CD4 T cells for your experiments. Sorting with this protocol gave us consistent, expected results.

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