Product nameAnti-SDHA antibody [2E3GC12FB2AE2]
See all SDHA primary antibodies
DescriptionMouse monoclonal [2E3GC12FB2AE2] to SDHA
Tested applicationsSuitable for: ICC, IHC-Fr, Flow Cyt, ICC/IF, WB, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Cow, Dog, Human
Full length native protein (purified). This information is considered to be commercially sensitive.
- Human heart mitochondria.
Storage instructionsShipped at 4°C. Store at +4°C.
Storage bufferPreservative: 0.02% Sodium azide
Constituent: HEPES buffered saline
Concentration information loading...
Purification notesNear homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
Light chain typekappa
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Integration of energy
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.
|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.
|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|
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.
- Information by UniProt
- CMD1GG antibody
- DHSA_HUMAN antibody
- Flavoprotein subunit of complex II antibody
All lanes : Anti-SDHA antibody [2E3GC12FB2AE2] (HRP) (ab198493) at 1/5000 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: 70 kDa
This data was developed using the same antibody clone in a different format (HRP conjugated) (ab198493).
ab198493 was shown to recognize in wild-type HEK-293 cells as signal was lost at the expected MW in SDHA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SDHA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab198493 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
250 μm-thick formalin-fixed cerebellum section were passively cleared using PACT and immunofluorescently labelled to identify mitochondrial mass (porin (ab14734, 1/100) and SDHA (ab14715, 1/100), 647 nm) and complex I subunits within the mitochondrial respiratory chain (NDUFB8 (ab110242, 1/100) and NDUFA13 (ab110240, 1/100); 546 nm) in conjunction with a neuronal marker (NF-H; 488 nm) in control 1. Scale: 100 μm.
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
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.
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).
All lanes :
Lanes 1 & 9 : Prestained Protein Ladder
Lanes 2-3 : Fly mitochondrial fraction
Lane 5 : Robin mitochondrial fraction
Lane 6 : Robin cytoplasmic fraction
Lane 7 : Mouse (AML12) mitochondrial fraction
Lane 8 : Mouse (AML12) cytoplasmic fraction
Predicted band size: 70 kDa
Mitochondrial and cytoplasmic fractions were obtained from whole flies with Abcam Mitochondrial Isolation Kit for Tissue (ab110168) and from robin fibroblast and mouse hepatocyte cell lines with Abcam Mitochondrial Isolation Kit for Cells (ab110170).
ab14715 was used to stain SDHA at 0.1 μg/ml
HRP-conjugated Horse Anti-mouse IgG at 1 μg/ml was used as a secondary antibody.
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 0.1% PBS-tween diluted 1%BSA (OR 10% goat serum OR 0.3M glycine) 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.
This product has been referenced in:
- Laurencé C et al. A new human pyridinium metabolite of furosemide, inhibitor of mitochondrial complex I, is a candidate inducer of neurodegeneration. Biochem Pharmacol 160:14-23 (2019). Read more (PubMed: 30537467) »
- Shin YS et al. Inhibition of bioenergetics provides novel insights into recruitment of PINK1-dependent neuronal mitophagy. J Neurochem N/A:N/A (2019). Read more (PubMed: 30664245) »