Clearly Shows Shift in CD154 Expression Among Peripheral Blood Leukocytes

Immunology
Duke University
Predoctoral

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Quality of Results

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Company:

eBioscience

Product Name:

Anti-Mouse CD154 (CD40 Ligand) APC

Catalog Number:

17-1541-82

The goal of this experiment was to phenotype CD154 expression among leukocytes in the peripheral blood of female C57Bl/6 mice.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Peripheral blood leukocytes

Primary Incubation

Stained cells in 100 uL cold PBS containing Live/Dead and TruStain Fc block (2 ug/mL) on ice for 15 minutes.

Blocking Agent

TruStain Fc block (2 ug/mL)

Secondary Incubation

Added 100 uL cold PBS containing isotype control (4 ug/mL) or CD154 APC(4 ug/mL) and incubated on ice for 15 minutes.

Tertiary Incubation

NA

Detection

FACS Canto

Results Summary

A clear shift in CD154 APC occurred for most peripheral blood leukocytes. A modest uptick in expression was observed for some cells, as noted by the doubling in the population contour.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Inexpensive antibody

The Bad

There wasn't a significant shift in the CD154+ leukocytes.

The Bottom Line

I would try a bright color conjugate, like PE, or a different clone.

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