Description
Total OXPHOS Rodent WB Antibody Cocktail ab110413 is an optimized cocktail of high quality antibodies for analyzing relative levels of OXPHOS complexes in rat or mouse mitochondria by western blot.
This OXPHOS cocktail 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 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).
Note: 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. 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.
The Western blot cocktail is supplied at a concentration of 1.5 mg/mL.
We recommend using a high pH CAPS / PVDF transfer protocol when using this antibody for Western blot.