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    products/primary-antibodies/sdhb-antibody-epr10880-ab175225.pdf

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Signal Transduction Metabolism Mitochondrial
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SDHB antibody [EPR10880] (ab175225)

  • Datasheet
  • SDS
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Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
  • Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunoprecipitation - Anti-SDHB antibody [EPR10880] (ab175225)
  • Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (ab175225)
  • Anti-SDHB antibody [EPR10880] (ab175225)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR10880] to SDHB
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Biotin Carrier Free HRP

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Overview

  • Product name

    Anti-SDHB antibody [EPR10880]
    See all SDHB primary antibodies
  • Description

    Rabbit monoclonal [EPR10880] to SDHB
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human SDHB aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P21912

  • Positive control

    • WB: HepG2, A431 and Jurkat whole cell lysate (ab7899) and human fetal heart, mouse brain and rat brain tissue lysates. IHC-P: Human caratoid paraganglioma, human mesenchymoma, human kidney, human liver, mouse kidney and rat kidney tissues. Flow Cyt (intra): Jurkat and A431 cells.
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR10880
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Oxidative phosphorylation
    • Complex II

Associated products

  • Alternative Versions

    • HRP Anti-SDHB antibody [EPR10880] (ab198329)
    • Biotin Anti-SDHB antibody [EPR10880] (ab198330)
    • Anti-SDHB antibody [EPR10880] - BSA and Azide free (ab249876)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human SDHB knockout HEK-293 cell line (ab260860)
  • KO cell lysates

    • Human SDHB knockout HEK-293 cell lysate (ab261652)
  • Positive Controls

    • Hep G2 whole cell lysate (ab166833)
    • Jurkat whole cell lysate (ab30128)
    • A-431 whole cell lysate (ab30132)
    • Mouse brain tissue lysate - total protein (ab30151)
    • Jurkat whole cell lysate (ab7899)
    • A-431 whole cell lysate (ab7909)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab175225 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/10 - 1/200.

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

WB
1/50000 - 1/200000. Predicted molecular weight: 32 kDa.
IP
1/10 - 1/100.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

Notes
Flow Cyt (Intra)
1/10 - 1/200.

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

WB
1/50000 - 1/200000. Predicted molecular weight: 32 kDa.
IP
1/10 - 1/100.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

Application notes
Is unsuitable for ICC/IF.

Target

  • Function

    Iron-sulfur protein (IP) 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).
  • Pathway

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

    Defects in SDHB are a cause of susceptibility to pheochromocytoma (PCC) [MIM:171300]. A catecholamine-producing tumor of chromaffin tissue of the adrenal medulla or sympathetic paraganglia. The cardinal symptom, reflecting the increased secretion of epinephrine and norepinephrine, is hypertension, which may be persistent or intermittent.
    Defects in SDHB are the cause of hereditary paragangliomas type 4 (PGL4) [MIM:115310]; also known as familial non-chromaffin paragangliomas type 4. Paragangliomas refer to rare and mostly benign tumors that arise from any component of the neuroendocrine system. PGL4 is characterized by the development of mostly benign, highly vascular, slow growing tumors in the head and neck. In the head and neck region, the carotid body is the largest of all paraganglia and is also the most common site of the tumors.
    Defects in SDHB are a cause of paraganglioma and gastric stromal sarcoma (PGGSS) [MIM:606864]; also called Carney-Stratakis syndrome. Gastrointestinal stromal tumors may be sporadic or inherited in an autosomal dominant manner, alone or as a component of a syndrome associated with other tumors, such as in the context of neurofibromatosis type 1 (NF1). Patients have both gastrointestinal stromal tumors and paragangliomas. Susceptibility to the tumors was inherited in an apparently autosomal dominant manner, with incomplete penetrance.
    Defects in SDHB are a cause of Cowden-like syndrome (CWDLS) [MIM:612359]. Cowden-like syndrome is a cancer predisposition syndrome associated with elevated risk for tumors of the breast, thyroid, kidney and uterus.
  • Sequence similarities

    Belongs to the succinate dehydrogenase/fumarate reductase iron-sulfur protein family.
    Contains 1 2Fe-2S ferredoxin-type domain.
    Contains 1 4Fe-4S ferredoxin-type domain.
  • Cellular localization

    Mitochondrion inner membrane.
  • Target information above from: UniProt accession P21912 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6390 Human
    • Entrez Gene: 67680 Mouse
    • Entrez Gene: 298596 Rat
    • Omim: 185470 Human
    • SwissProt: P21912 Human
    • SwissProt: Q9CQA3 Mouse
    • SwissProt: P21913 Rat
    • Unigene: 465924 Human
    • Unigene: 246965 Mouse
    • Unigene: 3902 Rat
    see all
  • Alternative names

    • CWS2 antibody
    • DHSB_HUMAN antibody
    • FLJ92337 antibody
    • Ip antibody
    • Iron sulfur subunit antibody
    • Iron sulfur subunit of complex II antibody
    • Iron-sulfur subunit of complex II antibody
    • mitochondrial antibody
    • PGL 4 antibody
    • PGL4 antibody
    • SDH 1 antibody
    • SDH antibody
    • SDH1 antibody
    • SDH2 antibody
    • SDH2, homolog of antibody
    • SdhB antibody
    • SDHIP antibody
    • Succinate dehydrogenase [ubiquinone] iron sulfur protein mitochondrial antibody
    • Succinate dehydrogenase [ubiquinone] iron sulfur subunit antibody
    • Succinate dehydrogenase [ubiquinone] iron-sulfur subunit antibody
    • succinate dehydrogenase [ubiquinone] iron-sulfur subunit, mitochondrial antibody
    • Succinate Dehydrogenase 1 Iron Sulfur Subunit antibody
    • Succinate Dehydrogenase 2, S. cerevisiae, homolog of antibody
    • Succinate dehydrogenase complex iron sulfur subunit B antibody
    • Succinate dehydrogenase complex subunit B iron sulfur antibody
    • Succinate Dehydrogenase Complex Subunit B Iron Sulfur Protein antibody
    • succinate dehydrogenase complex, subunit B, iron sulfur (Ip) antibody
    • Succinate dehydrogenase iron sulfur protein antibody
    see all

Images

  • Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
    Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
    All lanes : Anti-SDHB antibody [EPR10880] (ab175225) at 1/50000 dilution

    Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : SDHB knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 3 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 32 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab175225 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.

    ab175225 was shown to recognize SDHB in wild-type HEK 293 cells as signal was lost at the expected MW in SDHB knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SDHB knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab175225 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/50000 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-SDHB antibody [EPR10880] (ab175225)
    Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
    All lanes : Anti-SDHB antibody [EPR10880] (ab175225) at 1/100000 dilution (purified)

    Lane 1 : HepG2 whole cell lysate
    Lane 2 : Jurkat whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 32 kDa
    Observed band size: 32 kDa



    Blocking and dilution buffer: 5% NFDM/TBST
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labeling SDHB with unpurified ab175225 at a dilution of 1/100.

  • Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
    Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
    All lanes : Anti-SDHB antibody [EPR10880] (ab175225) at 1/100000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate
    Lane 3 : Mouse spleen tissue lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 32 kDa
    Observed band size: 32 kDa



    Blocking and dilution buffer: 5% NFDM/TBST
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human mesenchymoma tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling SDHB with purified ab175225 at a dilution of 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (ab175225)
    Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (ab175225)

    Intracellular Flow Cytometry analysis ofA431 cells labelling SDHB with purified ab175225 at a dilution of 1/200 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor®488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies. 

     

     

  • Immunoprecipitation - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunoprecipitation - Anti-SDHB antibody [EPR10880] (ab175225)

    ab175225 (purified) at a dilution of 1/60 immunoprecipitating SDHB in Jurkat whole cell lysate.

    Lane 1 (input): Jurkat whole cell lysate (10µg)

    Lane 2 (+): ab175225 + Jurkat whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab175225 in Jurkat whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
    Western blot - Anti-SDHB antibody [EPR10880] (ab175225)
    All lanes : Anti-SDHB antibody [EPR10880] (ab175225) at 1/50000 dilution (unpurified)

    Lane 1 : HepG2 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Fetal heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 32 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labeling SDHB with unpurified ab175225 at a dilution of 1/100.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHB antibody [EPR10880] (ab175225)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human carotid paraganglioma tissue labeling SDHB with unpurified ab175225 at a dilution of 1/100.

  • Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (ab175225)
    Flow Cytometry (Intracellular) - Anti-SDHB antibody [EPR10880] (ab175225)

    Intracellular flow cytometric analysis of permeabilized Jurkat cellslabeling SDHB with unpurified ab175225 at a dilution of 1/10 (red) compared toa rabbit IgG negative control (green). 

     

     

  • Anti-SDHB antibody [EPR10880] (ab175225)
    Anti-SDHB antibody [EPR10880] (ab175225)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab175225? Please let us know so that we can cite the reference in this datasheet.

ab175225 has been referenced in 2 publications.

  • Fang Z  et al. SDHB Suppresses the Tumorigenesis and Development of ccRCC by Inhibiting Glycolysis. Front Oncol 11:639408 (2021). PubMed: 34094922
  • Mot AI  et al. Circumventing the Crabtree Effect: A method to induce lactate consumption and increase oxidative phosphorylation in cell culture. Int J Biochem Cell Biol 79:128-138 (2016). PubMed: 27590850

Customer reviews and Q&As

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