Purification notesThe purity of ab110249 is near 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.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.25 µg/ml. Predicted molecular weight: 53 kDa.
Use a concentration of 5 µg/ml.
Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionCore subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I) that is believed to belong to the minimal assembly required for catalysis. Complex I functions in the transfer of electrons from NADH to the respiratory chain. The immediate electron acceptor for the enzyme is believed to be ubiquinone.
Involvement in diseaseDefects in NDUFS2 are a cause of mitochondrial complex I deficiency (MT-C1D) [MIM:252010]. A disorder of the mitochondrial respiratory chain that causes a wide range of clinical disorders, from lethal neonatal disease to adult-onset neurodegenerative disorders. Phenotypes include macrocephaly with progressive leukodystrophy, non-specific encephalopathy, cardiomyopathy, myopathy, liver disease, Leigh syndrome, Leber hereditary optic neuropathy, and some forms of Parkinson disease.
Sequence similaritiesBelongs to the complex I 49 kDa subunit family.
Overlay histogram showing HepG2 cells stained with ab110249 (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 followed by the antibody (ab110249, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Anti-NDUFS2 antibody [7A12BE5AD5] (ab110249)
All lanes : Anti-NDUFS2 antibody [7A12BE5AD5] (ab110249) at 0.25 µg/ml
Lane 1 : Molecular Weight Markers Lane 2 : Isolated mitochondria from Human liver Lane 3 : Isolated mitochondria from Bovine heart Lane 4 : Isolated mitochondria from H9C2 cells (Rat cardiomyocyte) Lane 5 : Isolated mitochondria from MEF cells (Mouse embryo fibroblast) Lane 6 : Isolated mitochondria from HepG2
Predicted band size : 53 kDa Complex I-subunit NDUFS2 is detected in the Human samples with Human specific ab110249, while Complex I-subunit NDUFB8 is detected in all samples (Human, Mouse and Rat) with an anti-NDUFB8.
Mitochondrial localization of Complex I subunit NDUFS2 visualized by Immunocytochemistry in HDFn cultured cells (normal Human dermal fibroblasts, neonatal), using ab110249 at 5 µg/ml.
References for Anti-NDUFS2 antibody [7A12BE5AD5] (ab110249)
This product has been referenced in:
Zurita Rendón O et al. A Mutation in the Flavin Adenine Dinucleotide-Dependent Oxidoreductase FOXRED1 Results in Cell-Type-Specific Assembly Defects in Oxidative Phosphorylation Complexes I and II. Mol Cell Biol36:2132-40 (2016).
Read more (PubMed: 27215383) »