使用敲除细胞株进行验证

Anti-NDUFA9抗体[20C11B11B11] (ab14713)

概述

  • 产品名称Anti-NDUFA9抗体[20C11B11B11]
    参阅全部 NDUFA9 一抗
  • 描述
    小鼠单克隆抗体[20C11B11B11] to NDUFA9
  • 经测试应用适用于: IHC-P, WB, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Cow, Human
  • 免疫原

    Tissue, cells or virus corresponding to Cow NDUFA9.

  • 阳性对照
    • 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.
  • 常规说明

    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.

性能

应用

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.

应用 Ab评论 说明
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 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

Anti-NDUFA9 antibody [20C11B11B11] 图像



  • Predicted band size : 40 kDa

    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 detected the expected band for NDUFA9 in wild-type HAP1 cells and the band was not seen in NDUFA9 knockout HAP1 cells. Wild-type and NDUFA9 knockout samples were subjected to SDS-PAGE. ab14713 and ab176560 (loading control to alpha tubulin) were diluted at 1 μg/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 1/10000 dilution for 1 hour at room temperature before imaging.

  • 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, 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.

  • 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
    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.
  • 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?)

Anti-NDUFA9 antibody [20C11B11B11] (ab14713)参考文献

This product has been referenced in:
  • Liu B  et al. Inhibition of ALDH2 by O-GlcNAcylation contributes to the hyperglycemic exacerbation of myocardial ischemia/reperfusion injury. Oncotarget 8:19413-19426 (2017). IP ; Rat . Read more (PubMed: 28038474) »
  • Gómez-Serrano M  et al. Differential proteomic and oxidative profiles unveil dysfunctional protein import to adipocyte mitochondria in obesity-associated aging and diabetes. Redox Biol 11:415-428 (2017). WB ; Human . Read more (PubMed: 28064117) »

See all 89 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Kidney)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Ventana CC1 (pH8.5)
Permeabilization No
Specification Kidney
Blocking step Ventana blocking agent as blocking agent for 4 minute(s) · Concentration: 100% · Temperature: 36°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Feb 23 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
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

提交于 Mar 12 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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

提交于 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
Abcam guarantees this product to work in the species/application used in this Abreview.
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

Brian Koss

Verified customer

提交于 Dec 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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

提交于 Nov 23 2011

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