Complex I Enzyme Activity Microplate Assay Kit (Colorimetric)

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PubMed

Functional Studies - Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) (AB109721)

Mitochondrial ROS dependent functional deficiency and structural impairment in cardiac mitochondria after sepsis.

Mitchondiral fractions from the heart tissue of Rats infected by S. pneumoniae, or given PBS sham control, were subjected to measurements of complex I-V activities. Complex I was measured with ab109721 (top left), Complex II + III were measured using ab109905 (top right), Complex IV was measured using ab109911 (bottom left) and Complex V was measured using ab109714 (bottom right).

Freshly isolated mitochondrial pellets were resuspened in PBS supplemented with 10% detergent provided in the kits. Protein concentrations of these mitochondrial lysates were estimated and 25 μg (for complex I, IV and V) or 100 μg (for complex II+III) mitochondrial protein was used per reaction. Enzyme activities were measured spectrophotometricly in triplicate and expressed as changes of absorbance per minute per mg protein.

Image courtesy of Yao X et al.PLoS One. 2015; 10(10): e0139416. doi: 10.1371/journal.pone.0139416.

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Supplier Data

Functional Studies - Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) (AB109721)

Principle of Complex I Enzyme Activity Microplate Assay Kit (ab109721)

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Functional Studies - Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) (AB109721)

ab109721 measures Complex I activity in human cultured cells within the recommended ranges given in the protocol. Example of Complex I activity measured in Hep2 cells is showed. (Note that these ranges depend on mitochondria preparation quality).

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Functional Studies - Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) (AB109721)

ab109721 measures Complex I activity in human and rat cultured cells but also in tissues/tissue mitochondria samples within the recommended ranges given in the protocol. Examples of Complex I activity measured in different rat tissue mitochondrial samples are shown. (Note that these ranges depend on mitochondria preparation quality).

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Supplier Data

Functional Studies - Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) (AB109721)

(A) ab109721 was used to measure Complex I activity in normal and Rho0 human fibroblast whole cell lysates at 1mg/mL. Rho0 cells are cells in which the mitochondrial DNA has been removed and therefore essential Complex I proteins are not expressed. As shown, the rho0 cells showed no/little complex I activity.

(B) In a similar analysis, rat cardiomyocytes were grown for 5 days in ± 40 μM chloramphenicol (CAM) to inhibit mitochondrial protein synthesis, Complex I assembly and hence activity was greatly reduced in samples loaded at 0.5 mg/mL whole cell lysates.

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Functional Studies - Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) (AB109721)

Examples of Complex I activity measured in different rat tissue mitochondrial samples are shown. (Note that these ranges depend on mitochondria preparation quality).