Contains 5 mAbs, one each against CI subunit NDUFB8 (MS105), CII-30kDa (MS203), CIII-Core protein 2 (MS304) CIV subunit I (MS404) and CV alpha subunit (MS507) 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.
Altered levels of 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 against a subunit that is labile when its complex is not assembled. Moreover, the combination is readily resolved in SDS-PAGE when the appropriate gel conditions are used (see protocols). Note: Mouse tissue samples can easily be contaminated with antibodies from the animal's blood. To avoid such background bands, use MS604 in conjunction with a secondary against native antibodies e.g. True blot (www.ebiosciences.com). Also, 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 loaded on the gel.