Hello, I am trying to isolate the mitochondria from brain tissue to see the effects of a certain drug administration on mitochondrial complexes and respiration. How can I extract the mitochondria without damaging it considering that initial steps of extraction involve tissue lysis by mechanical force with buffer (pulverization)?
Hi - MA Haidar. Yes, I agree with you that isolation of intact mitochondria is vital for undertaking the functional assays. In my Lab, we routinely use Mitochondria Isolation Kit from Thermo Scientific. I am providing you the link herewith:
https://assets.fishersci.com/TFS-Assets/LSG/manuals /MAN0011535_Mitochondria_Isolat_Tissue_UG.pdf
OR
https://www.biovision.com/documentation/datasheets/K288.pdf
Please follow - "Isolation of Mitochondria Using Reagent Based Method" given as an alternative (2nd Link - Pt. 2b).
Pradyumna Kumar Mishra thank you! So cutting the tissue, in the initial step of the tissue preparation, doesn't necessarily mean that the mitochondria will be affected due to mechanical force, right?
Yes, you are right. Gentle mincing of the tissue would not have any severe repercussion.
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