Hi, I used ExoQuick™Exosome Precipitation Solution to isolate exosomes from Plasma by following the standard protocol and resuspended the exosome pellet using 1X PBS buffer. I added NuPAGE™ LDS Sample Buffer (NP007) and NuPAGE Sample Reducing Agent (NP0009) to the solution and boiled at 90C for 5min to make sample for western blotting. However, the sample didn't run correctly. I found that two tracking dyes Coomassie G250 and Phenol Red didn't run in band shape, instead they lag behind compared with other samples and were smeared along the whole lane, some dyes still stack in wells.
Sheng Miao Do you think of washing your sample with PBS or lysate your exosome before running electrophoresis?
Nguyen Tien Cuong How to wash the sample with PBS but without losing exosome? I added NuPAGE™ LDS Sample Buffer (NP007) which contains detergent LDS to lysate exosome before running electrophoresis.
Sheng Miao A. Ahmed
I attached the method for isolation of exosome with Exoquick including washing with PBS in the paper in this comment.Lobb R. J. et al 2015 " Optimized exosome isolation protocol for cell culture supernatant and human plasma" Journal of extracellular vesicle. Article Optimized exosome isolation protocol for cell culture supern...
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