A fast, easy, cost-free method to remove excess dye or drug from exosome solution
Ioannis Isaioglou, Gloria Lopez-Madrigal, Jasmeen Merzaban
Disclaimer
The current protocol is now published in a peer-reviewed article at PLOS ONE doi.org/10.1371/journal.pone.0301761
Spotlight Video
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Abstract
This protocol details a cost-free method to remove excess dye or drug from exosome solution.
Attachments
Steps
Protocol Steps
Collect 0.5 x 106 cells (preferably parental cells used for exosome isolation)* and wash them with
phosphate buffered saline (PBS) or alternative buffer.
Pellet down the cells (typically 350x g).
Add the mixture containing the stained exosomes** with the excess dye/drug to the cell pellet.
Resuspend by gently pipetting the cells and leave the mixture shaking at 350rpm,37°C
Spin down the mixture so the cells form a pellet using the same parameters as in step 2.
Transfer the supernatant into a new clean tube. This supernatant should now contain the stained or loaded exosomes without any excess dye in the buffer solution used.
Repeat steps 2 through 5:
- Pellet down the cells (typically
350x g). - Add the mixture containing the stained exosomes** with the excess dye/drug to the cell pellet.
- Resuspend by gently pipetting the cells and leave the mixture shaking at
350rpm,37°C - Spin down the mixture so the cells form a pellet using the same parameters as in step 2.
Collect the supernatant which contains the stained exosomes without the excess dye.
Resuspend the pellet in PBS buffer (or alternative) and check whether the cells (test cells) were stained. This confirmation step can be verified using fluorescent microscopy or flow cytometry for fluorescent drugs or dyes.
