Anti-Complex I Immunocapture antibody [18G12BC2] (ab109798)
- Product nameAnti-Complex I Immunocapture antibody [18G12BC2]
- DescriptionMouse monoclonal [18G12BC2] to Complex I Immunocapture
- Tested applicationsIP, ICC, In-Cell ELISA, Flow Cyt more details
- Species reactivityReacts with: Mouse, Rat, Cow, Human
- Positive controlfibroblasts, HL-60 cells, tissue mitochondria
- Storage instructionsStore at +4°C. Do not freeze.
- Storage bufferPreservative: 0.02% Sodium azide
Constituent: 99% HBS
- Concentration information loading...
- PurityIgG fraction
- Purification notesNear homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
- Clonality Monoclonal
- Clone number18G12BC2
- Light chain typekappa
- Research Areas
Our Abpromise guarantee covers the use of ab109798 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||IP: Use at 100-1 µg/mg of lysate. 100 µg mAb can capture at least 25 µg complex I from 1 mg solubilized bovine heart mitochondria.|
|ICC||ICC: Use at an assay dependent dilution.|
|In-Cell ELISA||In-Cell ELISA: Use a concentration of 1 µg/ml.|
|Flow Cyt||Flow Cyt: Use 2µg for 106 cells.|
- RelevanceComplex I, or NADH ubiquinone oxidoreductase, is a large protein complex of 950,000 Da molecular weight made up by 45 to 46 different subunits. A total of seven of the subunits of the complex are encoded by mitochondrial DNA, while the remainder subunits are nuclear encoded, which are translated in the cytosol and translocated into the organelle for assembly at the inner membrane. The enzyme complex catalyses electron entry from NADH via a flavin (FMN) and several non-heme iron centers. Complex I is sensitive to a wide range of inhibitors, many of which are pesticides or other common environmental toxins, such as rotenone. Complex I dysfunction is a common cause of genetic OXPHOS defects. Altered functioning of this complex is also thought to contribute to several neurological disorders including Parkinson’s disease and schizophrenia. Also, there is evidence of Complex I involvement in diabetes.
Anti-Complex I Immunocapture antibody [18G12BC2] images
Complex I immunoprecipitation for antibody ab109798 crosslinked to protein G-agarose beads as product ab109711.
Immunocytochemistry image of ab109798 stained fibroblasts cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with the antibody (ab109798, 1 µg/mL) for 2 hours at room temperature or over night at 4°C. The secondary antibody was (green) Alexa Fluor® 4884 goat anti-mouse IgG (H+L) at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. The target protein locates to the mitochondria.
HL-60 cells were stained with 1 µg/mL Complex I antibody ab109798 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
Overlay histogram showing HepG2 cells stained with ab86750 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab867508, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-Complex I Immunocapture antibody [18G12BC2] (ab109798)
This product has been referenced in:
- Willis JH et al. Isolated deficiencies of OXPHOS complexes I and IV are identified accurately and quickly by simple enzyme activity immunocapture assays. Biochim Biophys Acta 1787:533-8 (2009). Read more (PubMed: 19041632) »
- Chinta SJ et al. Reactive oxygen species regulation by AIF- and complex I-depleted brain mitochondria. Free Radic Biol Med 46:939-47 (2009). Read more (PubMed: 19280713) »