Effective Antibody at Identifying Activated (CD80+) DCs

Cancer Center
George Washington University
Graduate Student

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Company:

BioLegend

Product Name:

PE/Cyanine7 anti-human CD80 Antibody

Catalog Number:

375408

Clone Number:

W17149D

I used this antibody to identify activated dendritic cell populations (CD80+) using flow cytometry, as part of a gating strategy, that involved gating on activated cells.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Human dendritic cells

Primary Incubation

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

Blocking Agent

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

Secondary Incubation

1uL of CD80 antibody diluted in 100uL of FACS buffer (5% FBS in DPBS), incubated at 4C for 20 mins

Tertiary Incubation

Cytofix buffer, 100uL for 20 mins at 4C

Detection

Flow cytometry in a beckman coulter cytoflex cytometer.

Results Summary

This product helped me identify activated dendritic cell populations in my cell culture protocol, which allows us to gate on dendritic cells (CD11c+) followed by CD80/CD86/CD40 for identification of activated dendritic cells. These results will be used to confirm DC activation during our maturation protocol and in the presence of activation-associated DAMPs such as LPS. It is important to note, due to my experimental design causing spectrums of activation, I did not get clearly positive and negative populations and therefore can't speak to the effectiveness of this antibody at identifying true negatives.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Able to identify activated (CD80+) dendritic cells.

The Bad

Spectrum of activation- no clearly identifiable positive and negative populations.

The Bottom Line

Antibody has great signal and is effective at identifying CD80+ cells.

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