Aldehyde/Sulfate Latex Beads for Exosome Flow Cytometry

August 31, 2018

Newcastle University
Academic Haematology
Research Associate

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Company:

ThermoFisher Scientific

Product Name:

Aldehyde/Sulfate Latex Beads 4% w/v

Catalog Number:

A37304

To allow detection of extracellular vesicles (EVs) in conventional cytometers, vesicles are coupled to 4 μm diameter aldehyde-sulfate latex beads. These can then be antibody labelled and applied to conventional flow cytometry protocols. We have used Aldehyde/sulphate latex beads 4% w/v in order to assist our flow cytometry protocol for detection of CD63 of EVs.

Experimental Design and Results Summary

Application

Flow cytometry analysis of EV CD63 expression

Starting Material

EVs isolated from cell conditioned media

Protocol Overview

We add 1ul of Aldehyde/sulphate latex beads to 4ul of EV suspension, and mix well. 1ml of particle-free PBS is then added, and the mixture left to incubate overnight at room temperature on a rotary wheel. 110ul of glycine is then added and the mixture incubated at room temperature for 30 minutes to block binding. The beads are then pelleted by centrifugation at 5,000RPM for 5 minutes, and the supernatant removed. The pellet is washed by addition of 1ml PBS+0.5% FBS and centrifugation at 5,000RPM for 5 minutes. The wash step is then repeated. Finally, the pellet is resuspended in PBS+0.5% FBS and antibodies of interest are added according to standard flow cytometry protocols.

Tips

Ensure careful removal of supernatant during the wash steps to minimise bead loss.

Results Summary

We get acceptable levels of positivity for EV markers according to flow cytometry results. This indicates effective binding of EVs to the Aldehyde/sulphate latex beads.

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DOI or PMID #

N/A

Additional Notes

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Summary

The Good

Simple to use and very small volumes required, ensuring the product goes a long way.

The Bad

Wash steps can be time consuming

The Bottom Line

An effective product to ensure ligation of EVs to latex beads in order to perform flow cytometry analysis.

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