Difference between revisions of "Template:SUIT text D066"
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::::The SUIT-029 O2 mt D066 is designed to evaluate the quality control of [[mitochondrial preparation]]. This protocol gives information of the linear coupling control ([[LEAK respiration|''L'']]-[[Oxidative phosphorylation| ''P'']]-[[ET-capacity| ''E'']]) with NADH linked-substrates ([[PM-pathway control state|PM]]). SUIT-029 O2 mt D066 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 preparation. When ATPases are active, residual endogenous adenylates are phosphorylated to ATP in the LEAK state, which is recycled to ADP in isolated mitochondria contaminated by ATPases, which leads to an overestimation of LEAK respiration, when comparing [[LEAK state without adenylates|''L(n)'']] and [[LEAK|''L(Omy)'']]. Therefore, it might be necessary to assess LEAK respiration in the presence of ATPase inhibitors, ''e.g.'' oligomycin. [[Additive effect of convergent electron flow]] through [[N-pathway control state|NADH-]] and [[NS-pathway control state| NS-pathways]] is evaluated in the [[ET-capacity| ET state]]. | ::::The SUIT-029 O2 mt D066 is designed to evaluate the quality control of [[mitochondrial preparation]]. This protocol gives information of the linear coupling control ([[LEAK respiration|''L'']]-[[Oxidative phosphorylation| ''P'']]-[[ET-capacity| ''E'']]) with NADH linked-substrates ([[PM-pathway control state|PM]]). SUIT-029 O2 mt D066 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 preparation. When ATPases are active, residual endogenous adenylates are phosphorylated to ATP in the LEAK state, which is recycled to ADP in isolated mitochondria contaminated by ATPases, which leads to an overestimation of LEAK respiration, when comparing [[LEAK state without adenylates|''L(n)'']] and [[LEAK respiration|''L(Omy)'']]. Therefore, it might be necessary to assess LEAK respiration in the presence of ATPase inhibitors, ''e.g.'' oligomycin. [[Additive effect of convergent electron flow]] through [[N-pathway control state|NADH-]] and [[NS-pathway control state| NS-pathways]] is evaluated in the [[ET-capacity| ET state]]. | ||
::::The present SUIT-029 O2 mt D066 DLP file shows an application with imt, thom or pce in the pathway category NS. For using this protocol with other mitochondrial preparations or substrate/inhibitor combinations, a personalized [[Export DL-Protocol User (*.DLPU)|DLPU]] can be created. | ::::The present SUIT-029 O2 mt D066 DLP file shows an application with imt, thom or pce in the pathway category NS. For using this protocol with other mitochondrial preparations or substrate/inhibitor combinations, a personalized [[Export DL-Protocol User (*.DLPU)|DLPU]] can be created. | ||
Revision as of 12:44, 25 November 2020
- The SUIT-029 O2 mt D066 is designed to evaluate the quality control of mitochondrial preparation. This protocol gives information of the linear coupling control (L- P- E) with NADH linked-substrates (PM). SUIT-029 O2 mt D066 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 preparation. When ATPases are active, residual endogenous adenylates are phosphorylated to ATP in the LEAK state, which is recycled to ADP in isolated mitochondria contaminated by ATPases, which leads to an overestimation of LEAK respiration, when comparing L(n) and L(Omy). Therefore, it might be necessary to assess LEAK respiration in the presence of ATPase inhibitors, e.g. oligomycin. Additive effect of convergent electron flow through NADH- and NS-pathways is evaluated in the ET state.
- The present SUIT-029 O2 mt D066 DLP file shows an application with imt, thom or pce in the pathway category NS. For using this protocol with other mitochondrial preparations or substrate/inhibitor combinations, a personalized DLPU can be created.
