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Negative Control for Exosomes

Hi,

  I have been purifying exosomes using the Total Exosome Isolation (from cell culture media) kit and am able to identify several exosomal markers by western blot (CD63, Alix, TSG1010, HSP90, Hsc70). However I recently also found Calnexin in my preparation (by western blot). Have you seen this before using this kit? What does it signify? Are there other markers that are better to use as "negative" controls for western blot?

Thank You.
asked Feb 13, 2015 in ExosomesTalk by asaxena

1 Answer

Hi,

 here is a few suggested by Lotvall et al in J of Extracellular vesicles (2014) which should be absent or under-represented in exosome preparations.

  • Endoplasmic reticulum
  • (Grp94HSP90B1,
  • calnexinCANX)
  • Golgi (GM130)
  • Mitochondria (cytochrome CCYC1)
  • Nucleus (histonesHIST*H*)
  • Argonaute/RISC complex (AGO*)

kind regards

ketil

answered Feb 16, 2015 by ketil.pedersen
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