Western Blots for Extracellular Vesicles: Tips, Tricks and Troubleshooting

Join our Senior Scientist Kate Timms (PhD) as she discusses Western blot analysis of extracellular vesicle (EV) samples. From experimental design and sample preparation, to lysis, loading, and blot evaluation, there are many aspects to consider. Understanding the potential pitfalls – and approaches to sidestepping them – can ensure your Western blot analysis of EVs post qEV isolation is as straightforward and thorough as possible.

00:00 Do Western blots require dark magic?
00:52 Considering controls to use in Western blots for EV analysis
01:30 Sample preparation and the importance of EV lysis
02:30 Determining the best conditions for EV sample lysis
03:25 How much sample to load? Should you compare by volume or by protein?
04:38 Reducing vs non-reducing loading buffer
05:49 Loading strategies and running your gel
06:30 Checking your gel after loading and considering further EV concentration
07:34 Detecting your protein of interest and antibody considerations
08:22 Interpreting your results

Related resources:
Read about Western blots for EV analysis: https://www.izon.com/news/a-step-by-s...

Learn about reproducible EV isolation using size exclusion chromatography: https://www.izon.com/qev/overview

Concentrate EVs quickly and cleanly using the qEV Magnetic Concentration Kit: https://www.izon.com/qev/kits