(MS601)
MitoProfile® Total OXPHOS Human WB Antibody Cocktail (ab110411)
Overview
- Product nameMitoProfile® Total OXPHOS Human WB Antibody Cocktail
- Assay typeQuantitative
- Species reactivityReacts with: Human
- Product overview
ab110411 (MS601) contains 5 mAbs, one each against Complex I subunit NDUFB8 (ab110242/MS105), Complex II subunit 30kDa (ab14714/MS203), Complex III subunit Core 2 (ab14745/MS304), Complex IV subunit II (ab110258/MS405), and ATP synthase subunit alpha (ab14748/MS507) as an optimized premixed cocktail. The cocktail is suitable for Western Blotting analysis of the relative levels of the 5 OXPHOS complexes in mitochondrial preparations from human sources. Human heart mitochondria (50ug) are also included as a Western Blotting control.
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).
Cocktail Antibodies:
Mouse monoclonal [20E9DH10C12] to (C-I-20) NDUFB8 (ab110242):
Amount: 50ug
Working concentration: 1 ug/mlMouse monoclonal [21A11AE7] to (C-II-30) SDHB (ab14714):
Amount: 250 ug
Working concentration: 5 ug/mlMouse monoclonal [13G12AF12BB11] to (C-III-Core 2) UQCRC2 (ab14745) :
Amount: 5 ug
Working concentration: 0.1 ug/mlMouse monoclonal [12C4F12] to (C-IV-II) Cytochrome C oxidase subunit II (ab110258):
Amount: 50ug
Working concentration: 1 ug/mlMouse monoclonal [15H4C4] to (C-V-alpha) ATP5A (ab14748):
Amount: 5 ug
Working concentration: 0.1 ug/ml - Notes
Store cocktail at 4°C and control at -80°C.
- Tested applicationsWB more details
Properties
- Storage instructionsStore at -80°C. Please refer to protocols.
-
Components 360 µg 720 µg Human Heart Mitochondria Western Blot Control 1 x 50µg 1 x 50µg Premix Cocktail of 5 monoclonal antibodies 1 x 360µg 1 x 720µg - Research Areas
-
Alternative names
- MS601-360OXPHOS cocktail
Applications
Our Abpromise guarantee covers the use of ab110411 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| WB | WB: Use a concentration of 7.2 µg/ml. The antibody cocktail (1.5 mg/mL) should be diluted 208x to a final working concentration of 7.2 µg/mL for Western blotting. |
MitoProfile® Total OXPHOS Human WB Antibody Cocktail images
-
Western Blot Abreview ImageThis image is courtesy of an anonymous AbreviewThis image is courtesy of an anonymous Abreview
Gel: Non-reduced Denaturing (4-12%)
Lanes 1-3: contain 3 individual Human B cell lymphoma samples 50 μg loaded per lane.
Blocked for 1 hour with 5% Milk at room temperature.
ab110411 used as per instructions.
Secondary: HRP-conjugated anti-mouse IgG uesd at 1/300
Detection method: ECL+ 2 minutes exposure.
-
Western blot - MitoProfile® Total OXPHOS Human WB Antibody Cocktail (ab110411)Each lane contains 10 µg of mitochondria isolated from MRC5 fibroblasts.
ab110411(MS601) was used to label lane 6, and the individual antibodies
comprising the cocktail were used for lanes 1-5.
References for MitoProfile® Total OXPHOS Human WB Antibody Cocktail (ab110411)
This product has been referenced in:
- Heo JY et al. DJ-1 null dopaminergic neuronal cells exhibit defects in mitochondrial function and structure: involvement of mitochondrial complex I assembly. PLoS One 7:e32629 (2012). WB . Read more (PubMed: 22403686) »
- Koob AO et al. Protein analysis through Western blot of cells excised individually from human brain and muscle tissue. Anal Biochem 425:120-124 (2012). WB . Read more (PubMed: 22402104) »
