Great CD31 Antibody for Staining Live Tissue Sections

July 26, 2018

BlueskyReddit
Developmental Immunology
La Jolla Institute for Allergy and Immunology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

BV421 anti-mouse CD31

Catalog Number:

102424

The goal of using this antibody was to stain the vasculature of mouse lung tissue. Lung tissue was embedded in agarose and sectioned into 300 micron sections using a vibratome. The sections were blocked for 1 hour with 24G2 Fc block on ice. Sections were then stained with the antibody at a 1:200 dilution overnight at 4 degrees Celsius. The sections were washed several times and fixed with 4% paraformaldehyde for 1 hour. Sections were washed several times and visualized by confocal microscopy.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Live, agarose-embedded mouse lung tissue sections

Primary Incubation

1:200 for 14 hours at 4 degrees celsius

Blocking Agent

3% BSA

Secondary Incubation

Fixed with 4% formaldehyde for 1 hour

Tertiary Incubation

none

Detection

Visualized by confocal microscopy

Results Summary

This antibody was highly specific for vascular endothelium in the lung sections. The staining was bright, and the antibody penetrated up to 150 microns into the tissue.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Very bright and specfic with low background

The Bad

BV421 is a large fluorophore, so signal was optimal at best up to 150 microns

The Bottom Line

This antibody stains live lung endothelial tissue with high specificity. It is also bright enough that a secondary antibody is not necessary for detection. I would recommend this antibody for this application.

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