Template:SUIT text D066

The SUIT-006 O2 mt D066 is designed to evaluate the quality control of mitochondrial preparation. This protocol includes a control for contamination with other organelles containing ATPases (e.g., plasma membrane and endoplasmic reticulum.) In experiments with crude isolated mitochondria,tissue homogenates and permeabilized cells, it is expected to have active ATPases in the prepation. When ATPases are active and residual endogenous adenylates are present in the sample, ATP can be consumed and converted to ADP, which will stimulate respiration. This may lead to an overestimation of LEAK respiration - L(n). Therefore, if the samples are contaminated with ATPases/adenylates, it might be necessary to assess LEAK respiration in the presence of ATPases inhibitors, e.g. oligomycin - L(Omy).

In the DatLab software, SUIT-006 O2 mt D066 DLP file is provided for mt, tissue homogenate or for cells which were permeabilized before addition into the O2k-Chamber and for the category NS. For using this protocol with other sample preparation or substrate/inhibitor combinations, a personalized DLPU can be created.