Product nameTotal OXPHOS Rodent WB Antibody Cocktail
Species reactivityReacts with: Mouse, Rat, Cow, Human, Cynomolgus monkey
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.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 300 µg Cocktail of 5 Antibodies 1 x 300µg Rat Heart Mitochondria Western Blot Control 1 x 50µg
- Anti-NDUFB8 antibody [20E9DH10C12] (ab110242)
- Anti-MTCO1 antibody [1D6E1A8] (ab14705)
- Anti-SDHB antibody [21A11AE7] (ab14714)
- Anti-UQCRC2 antibody [13G12AF12BB11] (ab14745)
- Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (ab14748)
- HeLa Mitochondrial DNA Depletion Cell Lysate Set: Ethidium Bromide-Treated and Vehicle-Treated (ab154479)
Our Abpromise guarantee covers the use of ab110413 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration.
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.
Store the antibody cocktail at 4°C and the control sample at -80°C.
We recommend using a high pH CAPS / PVDF transfer protocol when using this antibody for Western blot. Please contact us for more details.
Figure 1. Rat liver mitochondria labeled with ab110413 (MS604). The sample in lane 1 was kept at room temperature, whereas the remaining three samples were heated to 37°C, 50°C, and 100°C, respectively. This blot shows that boiling of samples leads to a decrease in signal due to aggregation of proteins therefore heating samples at or close to boiling is not recommended.
This product has been referenced in:
- Feng J et al. Simvastatin re-sensitizes hepatocellular carcinoma cells to sorafenib by inhibiting HIF-1a/PPAR-?/PKM2-mediated glycolysis. J Exp Clin Cancer Res 39:24 (2020). Read more (PubMed: 32000827) »
- Leermakers PA et al. Pulmonary inflammation-induced alterations in key regulators of mitophagy and mitochondrial biogenesis in murine skeletal muscle. BMC Pulm Med 20:20 (2020). Read more (PubMed: 31964384) »