Antibody from BioLegend Easily Identifies CD40+ Cells

George Washington University

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Brilliant Violet 421™ anti-human CD40 Antibody

Catalog Number:

334332

Clone Number:

5C3

The goal of this project was to identify activated cells in a CD11c+ and CD14+ (dendritic cell) primary human cell population after stimulation with an activation-inducing cytokine cocktail.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Human dendritic cells

Primary Incubation

0.1 uL Zombie Green viability stain in 100uL PBS, incubated at room temp for 15 mins.

Blocking Agent

Human TruStain Fc block, 5 uL diluted in 95 uL FACS buffer (5% FBS in DPBS), incubated at 4 degrees C for 10 mins

Secondary Incubation

1uL of CD40 antibody diluted in 100 uL of FACS buffer (5% FBS in DPBS), incubated at 4 degrees C for 20 mins.

Tertiary Incubation

Cytofix buffer, 100 uL for 20 mins at 4 degrees C.

Detection

Flow cytometry in a BD Celesta cytometer.

Results Summary

This product helped identify activated cells (CD40+ in combination with CD80/86) dendritic cells, to verify that our maturation and activation cocktails produce mature and activated dendritic cells. We were able to identify that almost all of our cells were CD40+.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Good identifier of CD40+ cells.

The Bad

Very bright, requires voltage adjustment.

The Bottom Line

Very bright antibody that identifies CD40+ cells.

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