Difference between revisions of "Template:SUIT text D056"
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SUIT-024 O2 ce-pce D056 is a protocol for permeabilized cells, designed to | SUIT-024 O2 ce-pce D056 is a protocol for permeabilized cells, designed to quantify the [[ATPases|ATPase]] activity in the sample. When [[Isolated mitochondria|isolating mitochondria]], contamination with other membranous organelles containing ATPases (''e.g.'', plasma membrane and endoplasmic reticulum) can occur. In experiments with tissue homogenates and permeabilized cells, it is expected to have active ATPases in the respiration medium. | ||
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 - [[LEAK state without adenylates|L(n)]]. Therefore, if the samples | 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 - [[LEAK state without adenylates|L(n)]]. Therefore, if the samples contains ATPases/adenylates, it might be necessary to assess LEAK respiration in the presence of ATPases inhibitors or oligomycin - [[LEAK|L(Omy)]]. This protocol can be used as a quality control of the [[mitochondrial preparation]]. | ||
In the DatLab software, SUIT-024 DLP file is provided for ce-pce and the category N(PM). For using this protocol with other sample preparation or substrate/inhibitor combinations, a personalized [[Export DL-Protocol User (*.DLPU)|DLPU]] can be created. | In the DatLab software, SUIT-024 DLP file is provided for ce-pce and the category N(PM). For using this protocol with other sample preparation or substrate/inhibitor combinations, a personalized [[Export DL-Protocol User (*.DLPU)|DLPU]] can be created. | ||
Revision as of 14:25, 16 April 2020
SUIT-024 O2 ce-pce D056 is a protocol for permeabilized cells, designed to quantify the ATPase activity in the sample. When isolating mitochondria, contamination with other membranous organelles containing ATPases (e.g., plasma membrane and endoplasmic reticulum) can occur. In experiments with tissue homogenates and permeabilized cells, it is expected to have active ATPases in the respiration medium. 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 contains ATPases/adenylates, it might be necessary to assess LEAK respiration in the presence of ATPases inhibitors or oligomycin - L(Omy). This protocol can be used as a quality control of the mitochondrial preparation.
In the DatLab software, SUIT-024 DLP file is provided for ce-pce and the category N(PM). For using this protocol with other sample preparation or substrate/inhibitor combinations, a personalized DLPU can be created.
