使用敲除细胞株进行验证重组RabMAb

重组Anti-SDHA抗体[EPR9043(B)] (ab137040)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9043(B)] to SDHA
  • Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

概述

  • 产品名称

    Anti-SDHA抗体[EPR9043(B)]
    参阅全部 SDHA 一抗

  • 描述

    兔单克隆抗体[EPR9043(B)] to SDHA

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human SDHA aa 550-650. The exact sequence is proprietary.

  • 阳性对照

    • WB: Wild-type HEK-293 whole cell lysate. MCF7, HT1080, Jurkat and HepG2 whole cell lysate. Mouse brain and kidney tissue lysate. Rat brain tissue lysate. IHC-P: Human, mouse and rat brain tissue. ICC: HeLa cells. IP: Jurkat and HeLa cell lysate. Flow Cyt: HeLa cells.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

性能

应用

Our Abpromise guarantee covers the use of ab137040 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
WB 1/1000 - 1/5000. Predicted molecular weight: 72 kDa.

Unpurified dilution 1/1000 - 1/10000.
IP 1/10 - 1/20.

Unpurified dilution 1/10 - 1/100.
Flow Cyt 1/10 - 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Unpurified dilution 1/10 - 1/100.
IHC-P 1/50 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.

靶标

  • 功能

    Flavoprotein (FP) subunit of succinate dehydrogenase (SDH) that is involved in complex II of the mitochondrial electron transport chain and is responsible for transferring electrons from succinate to ubiquinone (coenzyme Q).

  • 通路

    Carbohydrate metabolism; tricarboxylic acid cycle; fumarate from succinate (eukaryal route): step 1/1.

  • 疾病相关

    Defects in SDHA are a cause of mitochondrial complex II deficiency (MT-C2D) [MIM:252011]. A disorder of the mitochondrial respiratory chain with heterogeneous clinical manifestations. Clinical features include psychomotor regression in infants, poor growth with lack of speech development, severe spastic quadriplegia, dystonia, progressive leukoencephalopathy, muscle weakness, exercise intolerance, cardiomyopathy. Some patients manifest Leigh syndrome or Kearns-Sayre syndrome.
    Defects in SDHA are a cause of Leigh syndrome (LS) [MIM:256000]. LS is a severe disorder characterized by bilaterally symmetrical necrotic lesions in subcortical brain regions.
    Defects in SDHA are the cause of cardiomyopathy dilated type 1GG (CMD1GG) [MIM:613642]. CMD1GG is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.

  • 序列相似性

    Belongs to the FAD-dependent oxidoreductase 2 family. FRD/SDH subfamily.

  • 细胞定位

    Mitochondrion inner membrane.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • CMD1GG antibody
    • DHSA_HUMAN antibody
    • Flavoprotein subunit of complex II antibody
    • 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
    see all

图片

  • Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    All lanes : Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293 whole cell lysate
    Lane 2 : SDHA knockout HEK-293 whole cell lysate
    Lane 3 : MCF7 whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 72 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab137040 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab137040 was shown to specifically react with SDHA in wild-type HEK-293 cells as signal was lost in SDHA knockout cells. Wild-type and SDHA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab137040 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    All lanes : Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : HT1080 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP conjugated Goat anti Rabbit IgG at 1/2000 dilution

    Predicted band size: 72 kDa

  • Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    All lanes : Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/5000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 2 : HepG2 (human hepatocellular carcinoma) whole cell lysate
    Lane 3 : Jurkat (human acute T cell leukemia) whole cell lysate
    Lane 4 : Mouse brain tissue lysate
    Lane 5 : Mouse kidney tissue lysate
    Lane 6 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 72 kDa
    Additional bands at: 72 kDa. We are unsure as to the identity of these extra bands.



    Blocking buffer: 5% NFDM /TBST 

    Diluting buffer: 5% NFDM /TBST 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    ab137040 staining SDHA in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    ab137040 staining SDHA in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    ab137040 staining SDHA in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    Immunohistochemichal analysis of paraffin embedded human kidney tissue labelling SDHA with ab137040 at 1/50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    Immunohistochemichal analysis of paraffin embedded human testis tissue labelling SDHA with ab137040 at 1/50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling SDHA with ab137040 at 1/100 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    ab137040 staining SDHA in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab137040 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

  • Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    ab137040 immunoprecipitating SDHA. 10µg of Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: Jurkat whole cell lysate  10ug
    Lane 2: ab137040 IP in Jurkat whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab137040 in Jurkat whole cell lysate

  • Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    ab137040 immunoprecipitating SDHA. 10µg of HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: HeLa whole cell lysate (10ug)
    Lane 2: ab137040 IP in HeLa whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab137040 in HeLa whole cell lysate

  • Flow Cytometry - Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Flow Cytometry - Anti-SDHA antibody [EPR9043(B)] (ab137040)

    ab137040 staining SDHA in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/500 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Anti-SDHA antibody [EPR9043(B)] (ab137040)
    Anti-SDHA antibody [EPR9043(B)] (ab137040)

文献 (10)

发表研究结果有使用 ab137040？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab137040 被引用在 10 文献中.

  • Li B  et al. Mitochondrial-Derived Vesicles Protect Cardiomyocytes Against Hypoxic Damage. Front Cell Dev Biol 8:214 (2020). PubMed: 32426351
  • Liu Z  et al. IFI6 depletion inhibits esophageal squamous cell carcinoma progression through reactive oxygen species accumulation via mitochondrial dysfunction and endoplasmic reticulum stress. J Exp Clin Cancer Res 39:144 (2020). PubMed: 32727517
  • Liu L  et al. Exogenous nicotinamide adenine dinucleotide administration alleviates ischemia/reperfusion-induced oxidative injury in isolated rat hearts via Sirt5-SDH-succinate pathway. Eur J Pharmacol 858:172520 (2019). PubMed: 31278893
  • Meléndez-Rodríguez F  et al. HIF1a Suppresses Tumor Cell Proliferation through Inhibition of Aspartate Biosynthesis. Cell Rep 26:2257-2265.e4 (2019). PubMed: 30811976
  • Tang M  et al. Hippocampal proteomic changes of susceptibility and resilience to depression or anxiety in a rat model of chronic mild stress. Transl Psychiatry 9:260 (2019). PubMed: 31624233
  • Li H  et al. Iron regulatory protein deficiency compromises mitochondrial function in murine embryonic fibroblasts. Sci Rep 8:5118 (2018). PubMed: 29572489
  • Bao Y  et al. Carnosine Inhibits the Proliferation of Human Cervical Gland Carcinoma Cells Through Inhibiting Both Mitochondrial Bioenergetics and Glycolysis Pathways and Retarding Cell Cycle Progression. Integr Cancer Ther 17:80-91 (2018). PubMed: 28008780
  • Chen XF  et al. Effect of puerarin in promoting fatty acid oxidation by increasing mitochondrial oxidative capacity and biogenesis in skeletal muscle in diabetic rats. Nutr Diabetes 8:1 (2018). PubMed: 29330446
  • Riedl I  et al. AMPK?3 is dispensable for skeletal muscle hypertrophy induced by functional overload. Am J Physiol Endocrinol Metab 310:E461-72 (2016). PubMed: 26758685
  • Zecchini V  et al. Nuclear ARRB1 induces pseudohypoxia and cellular metabolism reprogramming in prostate cancer. EMBO J 33:1365-82 (2014). WB, ICC/IF . PubMed: 24837709

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1-3 of 3 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-SDHA antibody [EPR9043(B)]

 Excellent
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA
Permeabilization
No
Specification
brain
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

提交于 Oct 01 2019

Western blot abreview for Anti-SDHA antibody [EPR9043(B)]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Mitochondria isolated from mouse brain)
Gel Running Conditions
Reduced Denaturing (4-12% SDS-PAGE, Laemmli buffer with 10% β-Mercaptoethanol, 10 min at +55C denaturation. Immobilon-FL transfer membranes (Millipore), overnight transfer at +4C.)
Loading amount
15 µg
Specification
Mitochondria isolated from mouse brain
Blocking step
SEA Block Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C
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Abcam user community

Verified customer

提交于 Oct 17 2018

Western blot abreview for Anti-SDHA antibody [EPR9043(B)]

 Poor
Abreviews
abreview image
Application
Western blot
Loading amount
150 µg
Gel Running Conditions
Reduced Denaturing
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (Yeast whole cell)
Specification
Yeast whole cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Read More

Abcam user community

Verified customer

提交于 Dec 09 2013

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