Antibody titration for flow cytometric analysis of Extracellular Vesicles: Are cell based protocols up to the task?

Sponsored by: Miltenyi Biotec

Speaker

Event Date & Time

Date: October 6, 2021

Time: 9:00am (PDT), 12:00pm (EDT)

Abstract

ANTIBODY TITRATION is critical to the success of any flow cytometry experiment. This is even more so in microflow cytometric analysis of extracellular vesicles (EVs), where removing excess antibodies is technically challenging, if not practically impossible. In cell-based flow cytometry, improper dilution of an antibody is one of the main reasons for the lack of reproducible data. Too little antibody can lead to insufficient staining when excess antigen is present, while too much antibody can lead to antibody aggregation. Ideally, antibody concentration titration allows the separation of the positive signal from the negative background. While these caveats also apply to antibody titration for EVs, do cell-based protocols and analysis techniques translate directly to EV research?

In this webinar, we will investigate and report on antibody titration considerations for EVs, including sample concentration and processing, conjugate decisions, event triggering, platform differences, data processing and analysis, and a variety of real-world examples that highlight when antibody titration is necessary.

Learning Objectives:

Review the fundamentals of micro-flow cytometry (aka vesicle/high-resolution/nanoflow cytometry)
Discuss and understand the importance of antibody titration for flow cytometry experiments, concentrating on extracellular vesicle samples
Describe the criteria for choosing optimal antibody concentrations for EV experiments and how they may compare and differ from the criteria for choosing an optimal concentration for cell-based experiments

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Antibody titration for flow cytometric analysis of Extracellular Vesicles: Are cell based protocols up to the task?

Desmond Pink, PhD