Anti-NDUFA9 antibody [20C11B11B11]

• 产品名称

  Anti-NDUFA9抗体[20C11B11B11]

• 描述

  小鼠单克隆抗体[20C11B11B11] to NDUFA9

• 宿主

  Mouse

• 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.

  
  

  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

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C.

• 存储溶液

  pH: 7.4
  Preservative: 0.02% Sodium azide
  Constituent: HEPES buffered saline

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  IgG fraction

• 纯化说明

  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.

• 克隆

  单克隆

• 克隆编号

  20C11B11B11

• 同种型

  IgG1

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab14713于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

  hide

• SwissProt: Q5BK63 Rat

• Unigene: 75227 Human

• Unigene: 29939 Mouse

• Unigene: 107963 Rat

• 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

• 别名

• 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 39 kDa subunit antibody

• NDUA9_HUMAN antibody

• NDUFA9 antibody

• CI-39kD antibody

• Complex I subunit NDUFA9 antibody

• Complex I-39kD antibody

• 

  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.

• 

  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)

  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.

• 

  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 kDawhy 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)

  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 kDawhy is the actual band size different from the predicted?