a-CD39 with Exceptional Resolution

Protozoa Immunology
Bernhard Nocht Institut
Research Scientist

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Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

a-mouse CD39 PE-Cy7

Catalog Number:

143806

Clone Number:

Duha59

I used this antibody to analyze T cells in a “T cell-co-inhibitor/ exhaustion marker panel” in C57BL/6 mice infected with Plasmodium berghei ANKA, the murine model for malaria, in different organs.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

C57BL/6 splenocytes

Primary Incubation

1:200, 20 min, 4°C, in 50ul, in the dark

Blocking Agent

Fc Block

Secondary Incubation

Fix/Perm buffer, 20 min, 4°C, in 70ul, in the dark,

Tertiary Incubation

20 min, 4°C, in the dark, in 50µl Perm/wash

Detection

Flow cytomtetry

Results Summary

I used this antibody to analyze T cell regulation/ signs of exhaustion, in a murine model of malaria, in which C57BL/6 mice are infected with Plasmodium berghei ANKA. CD39, whilst not directly being an immune checkpoint, is often associated with T cell exhaustion This antibody gave me an exceptional CD39 signal, with an identifiable “positive” population. Often, markers in this category (PD-1, LAG-3,…) are seen as a “smear” rather than a distinct population; with this antibody, CD39 expression is clearly visible. At a titrated concentration of 1:200 (and still clearly detectable at lower concentrations), this antibody was easy to use for me whilst also being economical. This is my favourite CD39 clone.In the following, you will find my staining protocol in short. Incubation steps were performed at 4°C in the dark. 1.Count and adjust splenocytes to 1.5E6 cells, plate in a 96-well plate. 2.If required, stain cells with viability dye. Wash. 3. Stain with extracellular master mix for 20 min. Wash 2x. 4. Fixate/ Permeabilize with buffer of choice, depending on application. I used ThermoFisher FoxP3 fixation/permeabilization kit for 20 min. Wash with perm/ Wash 2x. 5. Perform intracellular staining for 20 min. Wash 2x. 6. Resuspend in FACS buffer and acquire as soon as possible. Here, data was acquired on a 5L Cytek Aurora or 5L BD LSRFortessa with the appropriate filter set.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Great resolution, even at lower concentrations.

The Bad

N/A

The Bottom Line

My favorite CD39 clone. Works well, great resolution. Strong signal even at a concentration of 1:800 (though I would recommend 1:200). Very economical in use.

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