Antibody From BioLegend Good for Showing a Shift but Not as Good at Showing Distinct CD83+ Population

September 22, 2023

George Washington University

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

PerCP/Cyanine5.5 anti-human CD83 Antibody

Catalog Number:

305320

Clone Number:

HB15e

The goal of my project was to visualize markers of dendritic cell activation, after gating on a CD11c+ population and inducing activation.

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 4 degrees C for 10 mins

Secondary Incubation

1uL of CD83 antibody diluted in 100uL of FACS buffer (5% FBS in DPBS), incubated at 4 degrees C for 20 mins

Tertiary Incubation

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

Detection

Flow cytometry in a BD Celesta cytometer

Results Summary

This antibody helped me visualize a shift in CD83 staining in activated and not activated conditions, but wasn't as good at giving a yes or no activation response due to the high overlap with a PE-tagged marker in the panel.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Stains CD83 on cells.

The Bad

High overlap with PE makes it difficult to find CD83 true positive population.

The Bottom Line

Antibody has good signal but you have to adjust voltage carefully and design panels carefully to avoid overlap, otherwise you're left with difficulty distinguishing positive populations from background/overlap.

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