Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
- Product nameAnti-NDUFA9 antibody [20C11B11B11]See all NDUFA9 primary antibodies ...
- DescriptionMouse monoclonal [20C11B11B11] to NDUFA9
- Tested applicationsIHC-P, WB, Flow Cyt more details
- Species reactivityReacts with: Mouse, Rat, Cow, Human
Purified mitochondrial complex I (Cow).
- Positive controlHuman heart mitochondria.
- Storage instructionsStore at +4°C.
- Storage bufferPreservative: 0.02% Sodium Azide
- 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 number20C11B11B11
- Light chain typekappa
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Integration of energy
Our Abpromise guarantee covers the use of ab14713 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||IHC-P: Use at an assay dependent concentration.|
|WB||WB: Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 40 kDa).|
|Flow Cyt||Flow Cyt: Use 1µg for 106 cells.|
- FunctionAccessory subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I), that is believed not to be involved in 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.
- Sequence similaritiesBelongs to the complex I NDUFA9 subunit family.
- Cellular localizationMitochondrion matrix.
- CI-39kD antibodyComplex I subunit NDUFA9 antibodyComplex I-39kD antibody
- mitochondrial antibodyNADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9 antibodyNADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9 mitochondrial antibodyNADH Ubiquinone Oxidoreductase 1 alpha subcomplex 9 antibodyNADH-Ubiquinone Oxidoreductase 1 alpha subcomplex 9 antibodyNADH-ubiquinone oxidoreductase 39 kDa subunit antibodyNDUA9_HUMAN antibodyNDUFA9 antibody
Anti-NDUFA9 antibody [20C11B11B11] images
Ab14713 staining human skeletal muscle. Staining is localized to the cytoplasm.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
All lanes : Anti-NDUFA9 antibody [20C11B11B11] (ab14713) at 1 µg/ml
Lane 1 : WI38 (Human lung fibroblast cell line) Whole Cell Lysate
Lane 2 :
Testis (Human) Tissue Lysate - adult normal tissue (ab30257)
Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 40 kDa
Observed band size : 36 kDa (why is the actual band size different from the predicted?)
Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 20 minutes
All lanes : Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
Lane 1 : Isolated mitochondria from Human heart at 5 µg
Lane 2 : Isolated mitochondria from Bovine heart at 1 µg
Lane 3 : Isolated mitochondria from Rat heart at 10 µg
Lane 4 : Isolated mitochondria from Mouse Heart at 10 µg
Goat anti-Mouse IgG
Predicted band size : 40 kDa
Observed band size : 37 kDa (why is the actual band size different from the predicted?)
ab14713 a 1/100 dilution staining NDUFA9 in Human spinal column tissue by Immunohistochemistry (Formalin/PFA-Fixed paraffin-embedded sections). Antibody was incubated with the sample for 1 hour. Sections were incubated in peroxidase-conjugated rabbit anti-mouse secondary (diluted 1/100 in 4% BSA in PBST) for 1 hour at room temperature. Sections were washed x3 in PBST and peroxidase activity was demonstrated using kit. Antigen retrieval was performed by 1 minute of pressure cooking with 1 mmol EDTA pH 8.0.
Overlay histogram showing HepG2 cells stained with ab14713 (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 (ab14713, 1µg/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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
This product has been referenced in:
- Kumarasamy S et al. Construction of two novel reciprocal conplastic rat strains and characterization of cardiac mitochondria. Am J Physiol Heart Circ Physiol 304:H22-32 (2013). WB ; Rat . Read more (PubMed: 23125210) »
- Yatsuga S & Suomalainen A Effect of bezafibrate treatment on late-onset mitochondrial myopathy in mice. Hum Mol Genet 21:526-35 (2012). Read more (PubMed: 22012983) »