Staining Monocytes with CX3CR1 PE

August 04, 2017

BlueskyReddit
Anaesthesiology
LMU Munich
Researcher

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Company:

Biolegend

Product Name:

PE anti-human CX3CR1 Antibody

Catalog Number:

355703

To identify Monocyte subpopulations involved in Monocyte adhesion andtransmigration, this antibody was used in a panel together with CD14, and CD16.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human Monocytes

Primary Incubation

10 mins room temperature (Fc Receptor blocking, 5µl for a 100µl cell suspension containing 2 milllion cells)

Blocking Agent

Tru Stain FcX (BioLegend), 10 Mins room temperature (5µl/100µl)

Secondary Incubation

2µl of antibody on 500000 cells, 30 mins on ice in the dark, wash three times afterwards with 2ml of FACS buffer (HBSS with 1%BSA and 0,01% Sodium Azide)

Tertiary Incubation

None

Detection

Flow Cytometry

Results Summary

This antibody allows for detection of CX3CR1 positive monocytes. Due to the fluorescent power of PE, even expressional changes in CX3CR1 surface expression may be identified using MFI calculations. After sufficient blocking, no unspecific staining was observed using a CX3CR1-negative subpopulation as a negative control. Results are reproducible using different lots of this antibody.Be sure to Fc-Block.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Easy to use, reliable, no background signal (after sufficient washing) or unspecific staining, reliable

The Bad

Blocking needs to be performed, extensice washing needed, pretty expensive antibody, PE is rather unstable.

The Bottom Line

Good antibody detecting also slight differences in CX3CR1 surface expression due PE brightness. Reliable, easy-to-use. Be aware of PE being a more unstable fluorophore than others.

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