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MitoSciences (MS204)

Anti-SDHA antibody [2E3GC12FB2AE2] - Mitochondrial Marker (ab14715)

Overview

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

    Purified mitochondrial complex II (Cow).

  • Positive controlHuman 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 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 IHC-Fr: Use at an assay dependent dilution.
Flow Cyt Flow Cyt: Use a concentration of 1 µg/ml.
WB WB: Use a concentration of 0.1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
IHC-P IHC-P: Use at an assay dependent dilution. 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 antibodyDHSA_HUMAN antibodyFlavoprotein subunit of complex II antibody
      Fp antibodyPGL5 antibodySDH 2 antibodySDH1 antibodySDH2 antibodySDHA antibodySDHF antibodySuccinate dehydrogenase [ubiquinone] flavoprotein subunit antibodySuccinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial antibodySuccinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial antibodySuccinate dehydrogenase complex flavoprotein subunit antibodySuccinate dehydrogenase complex flavoprotein subunit precursor antibodySuccinate dehydrogenase complex subunit A antibodySuccinate Dehydrogenase Complex subunit A Flavoprotein antibody
    see all

Anti-SDHA antibody [2E3GC12FB2AE2] - Mitochondrial Marker 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 antibody [2E3GC12FB2AE2] - Mitochondrial Marker (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 antibody [2E3GC12FB2AE2] - Mitochondrial Marker (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) »
  • Houstek J  et al. Nonsynonymous variants in mt-Nd2, mt-Nd4, and mt-Nd5 are linked to effects on oxidative phosphorylation and insulin sensitivity in rat conplastic strains. Physiol Genomics 44:487-94 (2012). WB ; Rat . Read more (PubMed: 22414913) »

See all 28 Publications for this product

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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

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Brain - Cerebellum)
Specification Brain - Cerebellum
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH8.0 EDTA
Permeabilization No
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Submitted Jun 28 2012

Application Western blot
Sample Mouse Cell lysate - other (Mouse Liver Mitochondria)
Loading amount 20 µg
Specification Mouse Liver Mitochondria
Gel Running Conditions Reduced Denaturing (10% Bis Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Dec 01 2011

Application Western blot
Sample Human Cell lysate - whole cell (HEK 293T)
Loading amount 20 µg
Specification HEK 293T
Gel Running Conditions Reduced Denaturing (10% Bis Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Dec 01 2011

Application Western blot
Sample Mouse Tissue lysate - other (mouse heart)
Loading amount 4 µg
Specification mouse heart
Gel Running Conditions Non-reduced Denaturing (10%)
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 07 2011

Application Western blot
Sample Hamster Cell lysate - other (mitochondria from Hamster CCL16 Cells (fibroblasts)
Loading amount 15 µg
Specification mitochondria from Hamster CCL16 Cells (fibroblasts
Gel Running Conditions Reduced Denaturing (14%)
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
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Submitted Jun 03 2008

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (293T)
Total protein in input 500 µg
Specification 293T
Immuno-precipitation step Protein G
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Submitted Oct 30 2007

Application Western blot
Sample Human Cell lysate - other (mitochondria from HeLa Cells)
Loading amount 15 µg
Specification mitochondria from HeLa Cells
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
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Submitted Oct 25 2007

Application Western blot
Sample Hamster Cell lysate - whole cell (CHO)
Loading amount 30 µg
Specification CHO
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Sep 17 2007

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (Cos-7 cell line)
Loading amount 30 µg
Specification Cos-7 cell line
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Sep 17 2007



Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"