Good Marker for Sorting CD4 T Cells

January 13, 2016

BlueskyReddit
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:

Pacific Blue™ Rat Anti-Mouse CD4

Catalog Number:

558107

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 as a marker for CD4+ 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, and CD44high/CD62Llow cells that we consider to be activated/memory T cells.

Additional Notes

None

Related Categories

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Summary

The Good

It was easy to identify the CD4+ T cells in total murine splenocytes.

The Bad

No drawbacks observed.

The Bottom Line

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

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