Hi, does anyone know the minimal criteria for Exosome marker? Because as far as I know, we can check CD9, CD63, CD81, HLA-G, Rab5b, Tsg 101 and Alix, but I can't find or not yet find the minimal criteria for exosome marker. And also do you know which best for the negative control?
Thank you & I really appreciate it.
Hi Arief,
CD8 marker labels all vacuolar compartments but it is not secreted as exosomes. It can be considered as a non-exosomal marker. You can get more information from this http://www.ncbi.nlm.nih.gov/pubmed/25154396 link
Try GM130, the cis-Golgi marker as a negative control. As for exosomal markers, I believe CD9 and CD63 would suffice, because they are more abundant in secreted exosomes compared, on a per mg of protein basis, to the total cell lysate.
You cannot find the minimum criteria because there is none agreed on 100%. It is conventionally agreed that complementary measures are necessary to claim you have exosomes. And that's still a claim depending on a definition. In the end you simply lay on the table various characterizations of your material and say: this is what WE call exosomes based on previous reports.
What you will most commonly find is an agreement of the following criteria taken together:
- size distribution with average 100-150 nm (not agreed on 100%) as detected by preferably both NTA and electron microscopy
- some exosomal marker and no golgi and ER marker (gm130, calnexin)
- bouyant density (from sucrose gradient or optiprep) - various reports, but you'll find a lot if you google this.
Cheers
Thank you very much for the answer, now I use CD9, CD63, TSG 101, ALIX, Rab5b and Flot-1 and for negative control I use Calnexin and GM130.
do you think its to much for Exosomes marker if I use 6 of them all together? I just thought, because TSG 101m ALIX and FLOT-1 is responsible for MVB or Biogenesis of Exosomes, so it would be indirect Quantification for Exosome. is there any comment on this particular issue?
thank you very much.
PS: I have trouble with my log in account, so it took a while for me to say "thank you"
The Journal of Extracellular Vesicles (JEV) recently published an article "Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles" (link below).
This should be able to clarify the minimum requirements for you.
Best of luck.
http://www.journalofextracellularvesicles.net/index.php/jev/article/view/26913
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