Invitrogen™ OxPhos Rodent WB Antibody Cocktail

Description

This product is a cocktail of multiple OxPhos antibodies. C-I-20 (ND6) (clone) 20E9DH10C12 - Bovine Complex I Purification Method: Antigen Affinity Chromatography C-II-30 (FeS) (clone) 21A11AE7 - Bovine Heart Complex II Purification Method: IgG Fraction C-III-Core 2 (clone) 13G12AF12BB11 - Bovine Heart Complex II, III Purification Method: IgG Fraction C-IV-I (clone) 1D6E1A8 - Human Complex IV subunit I Purification Method: Ammonium Sulfate Precipitation C-V-alpha (clone) 15H4C4 - Bovine Complex V Purification Method: Antigen Affinity Chromatography 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. These clones 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. 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 antibody cocktail (1.5 mg/mL) should be diluted 250x to a final working concentration of 6.0 μg/mL for Western blotting. We recommend using PBS with 1% milk as the antibody diluent and using a high pH CAPS/PVDF transfer protocol.

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.