Staining of CD40 as an Activation Marker on Murine Splenocytes

Tisch Cancer Institute
Icahn School of Medicine at Mount Sinai
Post-Doc

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

PE/Cy7 anti-mouse CD40 Antibody

Catalog Number:

124622

CD40 was stained on murine splenocytes to identify activated myeloid cells. Splenocytes were incubated with LPS overnight to induce activation of myeloid cells and then stained with CD11c and MHCII to identify dendritic cells. Anti-CD40 was used to measure activation of these cells. We chose PE-Cy7 due to its brightness to allow good discrimination between positive and negative cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine splenocytes

Primary Incubation

1:400, 15min, 4C

Blocking Agent

Blocking buffer containing sera (rat, rabbit, human, goat)

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow cytometry (BD) LSRFortessa

Results Summary

The antibody allows for easy identification of positive cells. Due to its brightness it can be used in relatively low concentrations.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Bright fluorophore

The Bad

Discrimination between positive and negative populations could be a little better

The Bottom Line

Easy to use, good staining quality, bright fluorophore

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