Different lanes represent different cell line derived exosome preparations
We used our patented technology which is soon to be launched in market. Exosomes were isolated from cell lines grown in exo-depleted FBS when they reached 80% confluency by collecting conditioned media.
Exosome yield depends on the amount of spent medium used for the isolation of exosomes. we have seen similar band intensity from exosomes prepared from 50 mL of spent medium from 1-2 million cells/mL.
Then they might not be bonafied exosomes. A mixture of apoptotic bodies, micro vesicles, and exosomes may result in this much protein. Have you subjected your preparation to size measurement?
When I say exosome preparations, inherently I mean that they have been characterized biophysically e.g. size, morphology (using DLS, SEM etc), as well as molecularly e.g. for exosomes marker proteins flotilin , TSG101 etc.
Well, then its fine. I am eager to try out this product once in the market.
Sure. Moreover, we have a kit (yet to be marketed) to extract maximum amount of proteins from exosome samples. I believe that also accounted for this much protein yield.
Hi Aman, At this size, I am afraid that it could be BSA which is a classical contaminant of exosome prep whatever the technique used tu purify them. Try an anti-BSA blot... I am also eager to try your product! good luck.
We have ruled out BSA presence as we run marker lane alongside which is shown in this CBB gel (the prominent band is above 120KD or so). Moreover, we use conditioned medium of 2 days (after cells reach 80% confluency; hence, devoid of any FBS if at all, and BSA therefore) Also, we have done some comparison with ultra exosome lysates which clearly shows CBB stained BSA band around 70 KD which our preparation does not show.
Before posting this question, we had done all sort of analysis, and ruled out various possibility frequently encountered by exosome researcher.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line