I am planning to isolate monocytes for Seahorse analysis and wondering wich isolation method to use,
and is the purity of the isolated monocytes of importance when doing Seahorse ?
Thank you for your question. The first thing I want to point out is that you want to make sure your cells are adhered to the bottom of the well. I am not sure which type of instrument you have, but below are our protocols for adhering non-adherent cells via Cell-Tak:
You do want them to be as pure as possible because any other cells that are seeded in the plate will contribute to your rates. We do not have an official recommendation as far as purifying methods, but you can use our Cell Reference Database to see what other users have done to purify. You can change the “Cell Type” drop-down to monocytes – there are many different papers listed that should be helpful!
Here are some papers I was able to find on there that I thought you might find interesting:
Additionally, here is a Jove Video specific to leukocytes that you may find helpful.
Please let me know if you have any other questions after taking a look at these documents/links.