What is the best way (fast) to disrupt macrophages in order to detect pro-il1b?
The best way to lyse cells are rapid freeze thaw. What you need to do is just dip the cells in liquid nitrogen and immediately thaw it in 37 degree water bath. Follow the procedure atleast 3 times immediately to get the cells lysed.
The best way to lyse cells are rapid freeze thaw. What you need to do is just dip the cells in liquid nitrogen and immediately thaw it in 37 degree water bath. Follow the procedure atleast 3 times immediately to get the cells lysed.
Thank you for the answer, but the problem is: The macrophages are attached to the plate. Is there a problem first disadhere, pass them to a eppendorff and only then do the procedure ?
Hi Aldo !! Vivek is right you can proceed in that way it's simple and in my case efficient. I used to do it with adherent cells as well, you must trypsin them first, but then you recuperate the cells and lyse them as Vivek said. In my case, I used to wash a lot after each step during the ELISA because of all the cellular debris that can disturb the test.
Best of luck.
Dear Aldo, Nadia is right. You can trypsin them or just do gentle pipetting with chilled PBS.
good luck.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA