MitoSciences (MS204)

Anti-SDHA [2E3GC12FB2AE2] antibody (ab14715)

Overview

  • Product nameAnti-SDHA [2E3GC12FB2AE2] antibodySee all SDHA primary antibodies ...
  • Description
    Mouse monoclonal [2E3GC12FB2AE2] to SDHA
  • Tested applicationsICC, IHC-Fr, Flow Cyt, ICC/IF, WB, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human, Caenorhabditis elegans
  • Immunogen

    Purified mitochondrial complex II (Cow).

  • Positive control
    • Human heart mitochondria.

Properties

Applications

Our Abpromise guarantee covers the use of ab14715 in the following tested applications.

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

Application Notes
ICC Use a concentration of 0.2 µg/ml. Requires heat-induced antigen retrieval where aldehydes are used as fixatives. Use 20min incubation at 90-100°C in 0.1 M Tris/HCl pH 9.5 with 5% urea (wt/vol).
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use a concentration of 1 µg/ml.
ICC/IF 1/200.
WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 20484225

Target

  • FunctionFlavoprotein (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).
  • PathwayCarbohydrate metabolism; tricarboxylic acid cycle; fumarate from succinate (eukaryal route): step 1/1.
  • Involvement in diseaseDefects 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.
  • Sequence similaritiesBelongs to the FAD-dependent oxidoreductase 2 family. FRD/SDH subfamily.
  • Cellular localizationMitochondrion inner membrane.
  • Target information above from: UniProt accession P31040 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • 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 antibody
    • Succinate dehydrogenase complex flavoprotein subunit precursor antibody
    • Succinate dehydrogenase complex subunit A antibody
    • Succinate Dehydrogenase Complex subunit A Flavoprotein antibody
    see all

Anti-SDHA [2E3GC12FB2AE2] antibody images

  • ICC/IF image of ab14715 stained human HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween 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.
  • 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.
  • All lanes : Anti-SDHA [2E3GC12FB2AE2] antibody (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 at 20 µg


    Predicted band size : 70 kDa
    Observed band size : 70 kDa
  • 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).
  • 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.
  • HL-60 cells were stained with 1 µg/mL ab14715 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.

References for Anti-SDHA [2E3GC12FB2AE2] antibody (ab14715)

This product has been referenced in:
  • Kumarasamy S  et al. Construction of two novel reciprocal conplastic rat strains and characterization of cardiac mitochondria. Am J Physiol Heart Circ Physiol 304:H22-32 (2013). WB ; Rat . Read more (PubMed: 23125210) »
  • Belinsky MG  et al. Succinate dehydrogenase deficiency in pediatric and adult gastrointestinal stromal tumors. Front Oncol 3:117 (2013). IHC-P ; Human . Read more (PubMed: 23730622) »

See all 39 Publications for this product

Product Wall

Thank you for contacting us. We do not know the epitopes to which this mAb binds. The immunogen was a full-length endogenous protein, of bovine origin; this antibody detects Hu, Ms, Rat, Ce, Cow, Dog SDHA protein and its specificity is fully guaran...

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Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Sample Human Cell (MDM)
Specification MDM
Permeabilization Yes - TRITON X
Fixative Paraformaldehyde
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Submitted Sep 03 2013

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - TRITON X
Sample Human Cell (MDM)
Specification MDM
Gating Strategy MACROPHAGE
Preparation Cell harvesting/tissue preparation method: SCRAPING
Sample buffer: PBS
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Submitted Sep 03 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Human Cell lysate - whole cell (MDM)
Specification MDM
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Sep 03 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (4-12)
Sample Rat Cell lysate - whole cell (Primary Neurons)
Specification Primary Neurons
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Jul 19 2013

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Sample Mouse Tissue sections (mouse heart)
Specification mouse heart
Permeabilization Yes - 1% triton x-100
Fixative Methanol
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Submitted Jul 02 2013

Western blot

Excellent
Abreviews
Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (hek293)
Specification hek293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted May 23 2013

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
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Submitted Mar 12 2013

Thank you for getting back to me and for providing some technical details.

This product has not been tested for IHC-P in our Lab. However, based on this specific publication (PubMed: 20484225) we can list it as tested application and offer ou...

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