Anti-SDHA antibody [2E3GC12FB2AE2]

• 产品名称

  Anti-SDHA抗体[2E3GC12FB2AE2]
  参阅全部 SDHA 一抗

• 描述

  小鼠单克隆抗体[2E3GC12FB2AE2] to SDHA

• 宿主

  Mouse

• 经测试应用

  适用于: IHC-Fr, Flow Cyt, WB, ICC, IHC-Pmore details

• 种属反应性

  与反应: Mouse, Rat, Cow, Human
  预测可用于: Dog

• 免疫原

  Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.

• 阳性对照

• WB: HEK-293, MCF7 amd HEepG2 whole cell lysates; Mitochondria isolated from human heart, cow heart, rat heart, mouse heart and HepG2 cells. Flow cyt: HL-60 cells. IHC-P: Human testis and skeletal muscle tissue. ICC: Cultured human embryonic lung-derived fibroblasts (strain MRC5).

• 常规说明

  
  

  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

  
  

  Product was previously marketed under the MitoSciences sub-brand.

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C.

• 存储溶液

  pH: 7.5
  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

  2E3GC12FB2AE2

• 同种型

  IgG1

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab14715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

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• SwissProt: Q8K2B3 Mouse

• SwissProt: Q920L2 Rat

• Unigene: 440475 Human

• Unigene: 158231 Mouse

• Unigene: 101725 Rat

• Entrez Gene: 281480 Cow

• Entrez Gene: 478634 Dog

• Entrez Gene: 6389 Human

• Entrez Gene: 66945 Mouse

• Entrez Gene: 157074 Rat

• Omim: 600857 Human

• SwissProt: P31039 Cow

• SwissProt: P31040 Human

• 别名

• Fp antibody

• PGL5 antibody

• SDH 2 antibody

• SDH1 antibody

• SDH2 antibody

• SDHA antibody

• SDHF antibody

• Succinate dehydrogenase [ubiquinone] flavoprotein subunit antibody

• Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial antibody

• Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial antibody

• Succinate dehydrogenase complex flavoprotein subunit A antibody

• Succinate dehydrogenase complex flavoprotein subunit antibody

• Succinate dehydrogenase complex flavoprotein subunit precursor antibody

• Succinate dehydrogenase complex subunit A antibody

• Succinate dehydrogenase complex subunit A flavoprotein (Fp) antibody

• Succinate dehydrogenase complex subunit A flavoprotein antibody

• CMD1GG antibody

• DHSA_HUMAN antibody

• Flavoprotein subunit of complex II antibody

• 

  Western blot - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)

  All lanes : HRP Anti-SDHA antibody [2E3GC12FB2AE2] (ab198493) at 1/5000 dilution
  
  Lane 1 : Wild-type HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
  Lane 2 : SDHA knockout HEK-293 whole cell lysate
  Lane 3 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
  Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Predicted band size: 70 kDa
  
  
  
  

  This data was developed using the same antibody clone in a different format (HRP conjugated) (ab198493).

  Lanes 1 - 4: Merged signal (red and green). Green - ab198493 observed at 72 kDa. Red - loading control, ab181602, observed at 37 kDa.

  ab198493 was shown to recognize in wild-type HEK-293 cells as signal was lost at the expected MW in SDHA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SDHA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab198493 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)Image from Phillips J et al., Sci Rep. 2016 May 16;6:26013. Fig 4a doi: 10.1038/srep26013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

  250 μm-thick formalin-fixed human cerebellum section were passively cleared using PACT and immunofluorescently labelled to identify mitochondrial mass (porin (ab14734, 1/100) and SDHA (ab14715, 1/100), 647 nm) and complex I subunits within the mitochondrial respiratory chain (NDUFB8 (ab110242, 1/100) and NDUFA13 (ab110240, 1/100); 546 nm) in conjunction with a neuronal marker (NF-H; 488 nm) in control 1. Scale: 100 μm.

• 

  Western blot - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)

  All lanes : Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)
  
  Lane 1 : Isolated mitochondria from Human heart at 5 µg
  Lane 2 : Isolated mitochondria from Bovine heart at 4 µg
  Lane 3 : Isolated mitochondria from Rat heart at 10 µg
  Lane 4 : Isolated mitochondria from Mouse heart at 10 µg
  Lane 5 : Isolated mitochondria from HepG2 (human liver hepatocellular carcinoma cell line) at 20 µg
  
  Predicted band size: 70 kDa
  Observed band size: 70 kDa
  
  

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)

  ab14715 (2µg/ml) staining SDHA in human testis using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic and mitochondrial staining within the seminal vesicles.
  Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  

• 

  Immunocytochemistry - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)

  Mitochondrial localization of complex II visualized by immunocytochemistry using ab14715. Cultured human embryonic lung-derived fibroblasts (strain MRC5) were fixed, permeabilized and then labeled with ab14715 (0.2 µg/ml) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG2a isotype specific secondary antibody (2 µg/ml).

• 

  Immunohistochemistry (Frozen sections) - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)

  Human skeletal muscle immunohistochemistry using ab14715. Fixed frozen tissue sections from a patient with a single large deletion of the mtDNA were used. All muscle fibers exhibit complex II immunoreactivity, consistent with the nuclear DNA-encoded expression pattern of this and all other subunits of complex II.

• 

  Immunocytochemistry - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)

  ICC/IF image of ab14715 stained human HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 0.1% PBS-tween diluted 1%BSA (OR 10% goat serum OR 0.3M glycine) for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14715, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in Hek293, HepG2 and MCF7 cells.

• 

  Flow Cytometry - Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)

  HL-60 (human promyelocytic leukemia cell line) cells were stained with 1 µg/mL ab14715 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.