Rat Anti-Mouse CD31 (Clone MEC13.3) Performs Well In Flow Cytometry

July 29, 2016

Division of BioMedical Sciences, Faculty of Medicine
Memorial University of Newfoundland
Research Assistant

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Alexa Fluor® 647 anti-mouse CD31 Antibody

Catalog Number:

102515

The general aim of our research is the study of lymphocyte biology and how this varies with health and disease. The specific goals of the experiments using this antibody were to monitor the levels of splenocyte CD31 expression in mice treated under different conditions. This antibody was chosen because (a) it is validated by the manufacturer for use in flow cytometry and (b) it is directly conjugated to a quality fluorophore.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse splenocytes

Primary Incubation

Alexa Fluor 647 anti-mouse CD31 antibody (BioLegend 102515), incubated with approximately 1x10^6 cells for 1 hour on ice at 1:200 dilution (0.25ug) in a total volume of 100ul PBS+2% FCS, 1mM EDTA, 0.1% sodium azide.

Blocking Agent

N/A

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

FACSCalibur (Becton Dickinson)

Results Summary

The majority of mouse splenocytes expressed CD31 (solid, open histogram) in comparison to cells incubated with an isotype control (broken, filled histogram). CD31-expressing cells were sufficiently bright enough to distinguish from the isotype control. Furthermore, a small proportion of mouse splenocytes clearly did not express CD31.

Additional Notes

None

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Summary

The Good

Little to no optimization required. Already labelled with a quality, trusted fluorophore.

The Bad

None

The Bottom Line

This product performed exactly as expected and reported by the manufacturer using their recommended protocol, antibody concentration, etc. Highly recommended for use in flow cytometry. Given that this antibody has been reported in the literature to work for IHC and IF, it would be helpful if the manufacturer quality tested this antibody for these applications.

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