Overview

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 Abreviews 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.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

  • Function
    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).
  • Pathway
    Carbohydrate metabolism; tricarboxylic acid cycle; fumarate from succinate (eukaryal route): step 1/1.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the FAD-dependent oxidoreductase 2 family. FRD/SDH subfamily.
  • Cellular localization
    Mitochondrion inner membrane.
  • 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 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

Images

  • 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] (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.
  • 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.

  • 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

This product has been referenced in:
  • Rohlenova K  et al. Selective Disruption of Respiratory Supercomplexes as a New Strategy to Suppress Her2(high) Breast Cancer. Antioxid Redox Signal 26:84-103 (2017). WB ; Human . Read more (PubMed: 27392540) »
  • Nguyen QL  et al. Platelets from pulmonary hypertension patients show increased mitochondrial reserve capacity. JCI Insight 2:e91415 (2017). Read more (PubMed: 28289721) »

See all 114 Publications for this product

Customer reviews and Q&As

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...

Read More
Application
Western blot
Sample
American Robin (Turdis migratorius) Cell lysate - other (fibroblasts, mitochondrial fraction)
Gel Running Conditions
Reduced Denaturing (18%)
Loading amount
25 µg
Specification
fibroblasts, mitochondrial fraction
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Sep 12 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - other (AML12 hepatocytes, mitochondrial fraction)
Gel Running Conditions
Reduced Denaturing (18)
Loading amount
25 µg
Specification
AML12 hepatocytes, mitochondrial fraction
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Aug 25 2017

Application
Western blot
Sample
Mouse Cell lysate - other (Mouse liver (A) and muscle (B) mitochondria)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-12%)
Loading amount
10 µg
Specification
Mouse liver (A) and muscle (B) mitochondria
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Jul 29 2015

Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Denaturing (14% 37.5:1)
Sample
Cow Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
3% Casein in PBS with 0.1% Tween-20 as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
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Submitted Feb 05 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Western blot

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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

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