Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
![Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)](https://www.abcam.com/ps/products/14/ab14713/Images/ab14713-268405-anti-ndufa9-antibody-20c11b11b11-western-blot.jpg "Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)")
Key features and details
- Mouse monoclonal [20C11B11B11] to NDUFA9
- Suitable for: IHC-P, WB, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Cow, Human
- Isotype: IgG1
Overview
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Product name
Anti-NDUFA9 antibody [20C11B11B11]
See all NDUFA9 primary antibodies -
Description
Mouse monoclonal [20C11B11B11] to NDUFA9 -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, WB, Flow Cytmore details -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human -
Immunogen
Tissue, cells or virus corresponding to Cow NDUFA9.
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Positive control
- WB: WI38 and NIH 3T3 whole cell lysates, human testis tissue lysate and human, cow, rat and mouse heart mitochondria. IHC-P: Human spinal column tissue. Flow Cyt: HepG2 cells.
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General notes
This monoclonal antibody to NDUFA9 has been knockout validated in Western blot. The expected band for NDUFA9 was observed in wild type cells and the band was not seen in knockout cells.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading... -
Purity
IgG fraction -
Purification notes
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. -
Clonality
Monoclonal -
Clone number
20C11B11B11 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P | (2) |
Use at an assay dependent concentration.
|
| WB | (5) |
Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 40 kDa).
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| Flow Cyt |
Use 1-2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
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IHC-P
Use at an assay dependent concentration. |
|
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 40 kDa). |
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Flow Cyt
Use 1-2µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Target
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Function
Accessory 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 similarities
Belongs to the complex I NDUFA9 subunit family. -
Cellular localization
Mitochondrion matrix. - Information by UniProt
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Database links
- Entrez Gene: 404188 Cow
- Entrez Gene: 4704 Human
- Entrez Gene: 66108 Mouse
- Entrez Gene: 362440 Rat
- Omim: 603834 Human
- SwissProt: P34943 Cow
- SwissProt: Q16795 Human
- SwissProt: Q9DC69 Mouse
see all -
Alternative names
- CI-39kD antibody
- Complex I subunit NDUFA9 antibody
- Complex I-39kD antibody
see all
Images
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Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: NDUFA9 knockout HAP1 cell lysate (20 µg)
Lane 3: WI38 cell lysate (20 µg)
Lane 4: NIH3T3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab14713 observed at 40 kDa. Red - loading control, ab176560, observed at 52 kDa.ab14713 was shown to specifically react with NDUFA9 in wild-type HAP1 cells. No band was observed when NDUFA9 knockout HAP1 samples were used. Wild-type and NDUFA9 knockout samples were subjected to SDS-PAGE. ab14713 and ab176560 (loading control to alpha tubulin) were diluted at 1μg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)](https://www.abcam.com/ps/products/14/ab14713/Images/ab14713-22045-anti-ndufa9-antibody-20c11b11b11-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)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. -
![Flow Cytometry - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)](https://www.abcam.com/ps/products/14/ab14713/Images/ab14713-22044-anti-ndufa9-antibody-20c11b11b11-flow-cytometry.jpg)
Flow Cytometry - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)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, 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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![Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)](https://www.abcam.com/ps/products/14/ab14713/Images/ab14713-22047-anti-ndufa9-antibody-20c11b11b11-western-blot.jpg)
Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)All lanes : Anti-NDUFA9 antibody [20C11B11B11] (ab14713) at 1 µg/ml
Lane 1 : WI38 (Human lung fibroblast cell line) Whole Cell Lysate
Lane 2 : Human testis tissue lysate - total protein (ab30257)
Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : 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
The band observed at 36 kDa could potentially be a cleaved form of NDUFA9 due to the presence of a 35 amino acid transit peptide. -
![Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)](https://www.abcam.com/ps/products/14/ab14713/Images/ab14713-22046-anti-ndufa9-antibody-20c11b11b11-western-blot.jpg)
Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)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
Secondary
All lanes : Goat anti-Mouse IgG
Predicted band size: 40 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Datasheets and documents
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SDS download
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Datasheet download
References (197)
ab14713 has been referenced in 197 publications.
- Ge M et al. APOL1 risk variants affect podocyte lipid homeostasis and energy production in focal segmental glomerulosclerosis. Hum Mol Genet 30:182-197 (2021). PubMed: 33517446
- Schober FA et al. The one-carbon pool controls mitochondrial energy metabolism via complex I and iron-sulfur clusters. Sci Adv 7:N/A (2021). PubMed: 33608280
- Todkar K et al. Selective packaging of mitochondrial proteins into extracellular vesicles prevents the release of mitochondrial DAMPs. Nat Commun 12:1971 (2021). PubMed: 33785738
- D'Angelo L et al. NDUFS3 depletion permits complex I maturation and reveals TMEM126A/OPA7 as an assembly factor binding the ND4-module intermediate. Cell Rep 35:109002 (2021). PubMed: 33882309
- Silwal P et al. Mitofusin-2 boosts innate immunity through the maintenance of aerobic glycolysis and activation of xenophagy in mice. Commun Biol 4:548 (2021). PubMed: 33972668