Great CD11c+ Marker From BioLegend

August 20, 2023

BlueskyReddit
George Washington University

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Quality of Results

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Company:

BioLegend

Product Name:

Alexa Fluor® 700 anti-human CD11c Antibody

Catalog Number:

337220

Clone Number:

Bu15

The goal of this project was to mature CD14+ PBMCs (monocytes) into dendritic cells, then analyze activation in the presence of activation-inducing cytokines.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Human dendritic cells

Primary Incubation

0.1 uL Zombie Green viability stain in 100 uL 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

2 uL of CD11c 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 product helped me gate on CD11c+ cells to differentiate true dendritic cells from other monocyte-derived cells or PBMCs (if they have survived). I identified close to 100% CD11c+ cells, which was what I expected to find given that I isolated for monocytes and cultured them with dendritic cell maturation cytokines.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Identifies CD11c+ cells.

The Bad

Needed 2 uL to show clear separation of negative and positive when titrating.

The Bottom Line

Not too expensive and works as a good marker to establish dendritic cell population (after gating on or isolating CD14+ cells).

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