MitoSciences (MS111)

Anti-NDUFA9 antibody [20C11B11B11] (ab14713)

Overview

  • Product nameAnti-NDUFA9 antibody [20C11B11B11]See all NDUFA9 primary antibodies ...
  • Description
    Mouse monoclonal [20C11B11B11] to NDUFA9
  • Tested applicationsIHC-P, WB, Flow Cyt more details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Purified mitochondrial complex I (Cow).

  • Positive control
    • Human heart mitochondria.

Properties

Applications

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 Notes
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).
Flow Cyt Use 1µg for 106 cells.

Target

  • 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.
  • Target information above from: UniProt accession Q16795 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • CI-39kD antibody
    • Complex I subunit NDUFA9 antibody
    • Complex I-39kD antibody
    • mitochondrial antibody
    • NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9 antibody
    • NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9 mitochondrial antibody
    • NADH Ubiquinone Oxidoreductase 1 alpha subcomplex 9 antibody
    • NADH-Ubiquinone Oxidoreductase 1 alpha subcomplex 9 antibody
    • NADH-ubiquinone oxidoreductase 39 kDa subunit antibody
    • NDUA9_HUMAN antibody
    • NDUFA9 antibody
    see all

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.

    Secondary
    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

    Secondary
    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:
  • Davoudi M  et al. Complex I function and supercomplex formation are preserved in liver mitochondria despite progressive complex III deficiency. PLoS One 9:e86767 (2014). WB ; Mouse . Read more (PubMed: 24466228) »
  • 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) »

See all 67 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (hek293)
Specification hek293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

Submitted Jul 23 2013

Application Western blot
Sample Rat Cell lysate - whole cell (h9c2)
Loading amount 30 µg
Specification h9c2
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

Submitted Mar 12 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (6-week old testes)
Specification 6-week old testes
Fixative 4% PFA
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 0.01M citric acid/citrate sodium
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Qing Wen

Verified customer

Submitted Jan 03 2013

Thank you for your enquiry.

I can confirm that the following antibodies are produced in our Mitosciences laboratories:


ab14713
ab110244
ab14715
ab14745
ab14705

These have been tested in blue nat...

Read More
Application Western blot
Sample Human Cell lysate - whole cell (HEK 293T)
Loading amount 20 µg
Specification HEK 293T
Gel Running Conditions Reduced Denaturing (10% Bis Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Brian Koss

Verified customer

Submitted Dec 01 2011

Application Western blot
Sample Mouse Tissue lysate - other (Liver Mitochondria)
Loading amount 20 µg
Specification Liver Mitochondria
Gel Running Conditions Reduced Denaturing (10% Bis Trix)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Nov 23 2011

Application Western blot
Sample Mouse Tissue lysate - other (mouse heart)
Loading amount 4 µg
Specification mouse heart
Gel Running Conditions Non-reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 22 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"