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:
Immobilization of Non-Adherent Cells with Cell-Tak for Assay on the Agilent Seahorse XFe/XF24 Analyzer
Immobilization of Non-Adherent Cells with Cell-Tak for Assay on the Agilent Seahorse XFe/XF96 or XFp Analyzer
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:
1. Rewiring monocyte glucose metabolism via C-type lectin signaling protects against disseminated candidiasis
11. Assessing Functional Metabolism Using Cells Sourced from Blood Real-time, Kinetic Measurements Link Mitochondrial Function to Disease Diagnosis and Progression
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.
This post appears to be related to our Seahorse products, which unfortunately are not currently supported in this community. The best way for you to get help with this is to email email@example.com.
Thank you for your reply, I will send an email.
I have similar query and have been struggling to get any attention after emailing firstname.lastname@example.org.
Is there any better way of getting through to support/advice?
Hi simao86, we will have our support team reach out to you for follow up.
They have thank you
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