Total OXPHOS Rodent WB Antibody Cocktail(AB110413)

Total OXPHOS Rodent WB Antibody Cocktail Details 

This OXPHOS cocktail ab110413 contains 5 mouse mAbs, one each against CI subunit NDUFB8 (ab110242), CII-30kDa (ab14714), CIII-Core protein 2 (ab14745), CIV subunit I (ab14705) and CV alpha subunit (ab14748) as an optimized premixed cocktail. The kit is suitable for Western Blotting analysis of the relative levels of the 5 OXPHOS complexes in mitochondrial preparations from mouse, rat, human, or bovine sources. The positive control supplied with the cocktail is ab110341. 

Altered levels of OXPHOS complex assembly can arise from mutations in individual subunits, mutations in assembly factors for the complex(es), mtDNA depletion or as a result of physiological and or pathological changes e.g. hormone treatment, exercise, diet or oxidative stress. 

The mAbs in the cocktail were chosen because they are each against a subunit that is labile when its complex is not assembled. Also, the different subunits are easily resolved in SDS-PAGE (see protocols).

Tips for success 

Mouse tissue samples can easily be contaminated with antibodies from the animal's blood. To avoid background bands, use ab110413 with an anti-mouse secondary against native antibodies. We recommend Anti-mouse IgG for IP (HRP) ab131368 which detects native mouse IgG antibodies (subtypes: IgG1, IgG2a, IgG2b, IgG3). 

COXI is a highly hydrophobic protein and appears as a broad band at ~35 kDa (not at its true molecular weight at 57 kDa). It is very sensitive to heating. Therefore, the samples, including the positive control, should not be heated over 50°C before loading on the gel. 

The antibody cocktail (1.5 mg/mL) should be diluted 250x to a nal working concentration of 6.0 µg/mL for Western blotting. 

We recommend using PBS with 1% milk as the antibody diluent, and a high pH CAPS / PVDF transfer protocol. Please contact us for more details. 

The Western blot cocktail is supplied at a concentration of 1.5 mg/mL. Store the antibody cocktail at 4°C and the control sample at -80°C.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

MitoProfile^® Total OXPHOS Rodent refers to a collection of proteins associated with oxidative phosphorylation (OXPHOS) in rodents. This target includes several key protein complexes involved in the electron transport chain within mitochondria. These complexes specified as OXPHOS Complexes I-V act in conjunction to facilitate ATP production. OXPHOS proteins such as NADH dehydrogenase (complex I) cytochrome c oxidase (complex IV) and ATP synthase (complex V) are detected using antibodies specified in a mitochondrial cocktail or OXPHOS cocktail tailored for Western blots. Mitochondria express these proteins across various tissues including skeletal muscle heart and brain reflecting their essential role in energy metabolism.

Biological function summary

OXPHOS proteins are integral to the production of ATP the primary energy molecule in cells. They form a multi-protein complex that transduces energy by transferring electrons and pumping protons across the inner mitochondrial membrane. The OXPHOS system comprises complexes I to V each playing a specific role within this energy-converting process. This system is important for cellular respiration acting as the powerhouse that drives numerous cellular processes.

Pathways

OXPHOS proteins participate significantly in the mitochondrial electron transport chain and oxidative phosphorylation pathways. The ATP produced through this mechanism contributes to various metabolic pathways and cellular activities. Cytochrome c associated with complex IV plays a pivotal role in apoptotic pathways linking energy metabolism with cell death processes. Other key proteins such as ubiquinone function as electron carriers in the electron transport chain to ensure efficient ATP synthesis.

Impaired OXPHOS function associates with metabolic disorders such as mitochondrial diseases and neurodegenerative conditions like Parkinson's disease. Defects in any of the OXPHOS complexes can lead to insufficient ATP production causing energy deficits in cells and tissues. For example complex I defects can contribute to degenerative disorders linked with lease syndrome through dysregulated energy metabolism. Variations in expression of components within the OXPHOS system such as mutations or deletions can also influence the risk and progression of these diseases.

Target data

Accessory subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I), that is believed not to be involved in catalysis. Complex I functions in the transfer of electrons from NADH to the respiratory chain. The immediate electron acceptor for the enzyme is believed to be ubiquinone.

Sample Images for Expected Results & Reference