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.

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 was shown to specifically react with NDUFA9 when NDUFA9 knockout 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 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:
  • Sakakibara I  et al. Six1 homeoprotein drives myofiber type IIA specialization in soleus muscle. Skelet Muscle 6:30 (2016). WB ; Mouse . Read more (PubMed: 27597886) »
  • Pérez-Pérez R  et al. COX7A2L Is a Mitochondrial Complex III Binding Protein that Stabilizes the III2+IV Supercomplex without Affecting Respirasome Formation. Cell Rep 16:2387-98 (2016). WB . Read more (PubMed: 27545886) »

See all 83 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"