Invitrogen™ OxPhos Rodent WB Antibody Cocktail

Beschreibung

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 50ºC 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.