Name: Anti-SDHA antibody [EPR9043(B)]
Brand: RabMAb
SKU: ab137040
Rating: 4 (4 reviews)

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Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR9043(B)] to SDHA
Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-SDHA antibody [EPR9043(B)]
See all SDHA primary antibodies
Description

Rabbit monoclonal [EPR9043(B)] to SDHA
Host species

Rabbit
Tested applications

Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide within Human SDHA aa 550-650. The exact sequence is proprietary.
Positive control
- WB: Wild-type HEK-293 whole cell lysate. MCF7, HT1080, Jurkat and HepG2 whole cell lysate. Mouse brain and kidney tissue lysate. Rat brain tissue lysate. IHC-P: Human, mouse and rat brain tissue. ICC: HeLa cells. IP: Jurkat and HeLa cell lysate. Flow Cyt (intra): HeLa 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.

Form

Liquid
Storage instructions

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

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR9043(B)
Isotype

IgG
Research areas

Alternative Versions
- Anti-SDHA antibody [EPR9043(B)] - BSA and Azide free (ab240098)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human SDHA knockout HEK-293 cell line (ab261853)
KO cell lysates
- Human SDHA knockout HEK-293 cell lysate (ab261657)
Positive Controls
- HeLa whole cell lysate (ab29545)
- Jurkat whole cell lysate (ab7899)
Recombinant Protein
- Recombinant Human SDHA protein (Tagged) (ab226268)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab137040 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
WB	(2)	1/1000 - 1/5000. Predicted molecular weight: 72 kDa. Unpurified dilution 1/1000 - 1/10000.
IP		1/10 - 1/20. Unpurified dilution 1/10 - 1/100.
IHC-P	(1)	1/50 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF		1/100 - 1/250.
Flow Cyt (Intra)		1/10 - 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Unpurified dilution 1/10 - 1/100.

Notes
WB 1/1000 - 1/5000. Predicted molecular weight: 72 kDa. Unpurified dilution 1/1000 - 1/10000.
IP 1/10 - 1/20. Unpurified dilution 1/10 - 1/100.
IHC-P 1/50 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
Flow Cyt (Intra) 1/10 - 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Unpurified dilution 1/10 - 1/100.

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.
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
- Entrez Gene: 6389 Human
- Entrez Gene: 66945 Mouse
- Entrez Gene: 157074 Rat
- Omim: 600857 Human
- SwissProt: P31040 Human
- SwissProt: Q8K2B3 Mouse
- SwissProt: Q920L2 Rat
- Unigene: 440475 Human
- Unigene: 158231 Mouse
- Unigene: 101725 Rat
see all
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

Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)

All lanes : Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/1000 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: 72 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab137040 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab137040 was shown to specifically react with SDHA in wild-type HEK-293 cells as signal was lost in SDHA knockout cells. Wild-type and SDHA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab137040 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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-SDHA antibody [EPR9043(B)] (ab137040)

All lanes : Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/1000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : HT1080 cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP conjugated Goat anti Rabbit IgG at 1/2000 dilution

Predicted band size: 72 kDa
Western blot - Anti-SDHA antibody [EPR9043(B)] (ab137040)

All lanes : Anti-SDHA antibody [EPR9043(B)] (ab137040) at 1/5000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2 : HepG2 (human hepatocellular carcinoma) whole cell lysate
Lane 3 : Jurkat (human acute T cell leukemia) whole cell lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : Mouse kidney tissue lysate
Lane 6 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 72 kDa
Additional bands at: 72 kDa. We are unsure as to the identity of these extra bands.

Blocking buffer: 5% NFDM /TBST

Diluting buffer: 5% NFDM /TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

ab137040 staining SDHA in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1: PBS in place of primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

ab137040 staining SDHA in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1: PBS in place of primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

ab137040 staining SDHA in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/1000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1: PBS in place of primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

Immunohistochemichal analysis of paraffin embedded human kidney tissue labelling SDHA with ab137040 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

Immunohistochemichal analysis of paraffin embedded human testis tissue labelling SDHA with ab137040 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (ab137040)

Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling SDHA with ab137040 at 1/100 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-SDHA antibody [EPR9043(B)] (ab137040)

ab137040 staining SDHA in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab137040 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)
Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)

ab137040 immunoprecipitating SDHA. 10µg of Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

Lane 1: Jurkat whole cell lysate 10ug
Lane 2: ab137040 IP in Jurkat whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab137040 in Jurkat whole cell lysate
Immunoprecipitation - Anti-SDHA antibody [EPR9043(B)] (ab137040)

ab137040 immunoprecipitating SDHA. 10µg of HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

Lane 1: HeLa whole cell lysate (10ug)
Lane 2: ab137040 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab137040 in HeLa whole cell lysate
Flow Cytometry (Intracellular) - Anti-SDHA antibody [EPR9043(B)] (ab137040)

ab137040 staining SDHAin the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/500 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Anti-SDHA antibody [EPR9043(B)] (ab137040)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab137040? Please let us know so that we can cite the reference in this datasheet.

ab137040 has been referenced in 14 publications.

Xu X et al. Upregulation of SDHA inhibited proliferation, migration, and invasion of clear cell renal cell carcinoma cells via inactivation of the Wnt/ß-catenin pathway. J Recept Signal Transduct Res N/A:1-13 (2021). PubMed: 33602019
Wang Z et al. High throughput proteomic and metabolic profiling identified target correction of metabolic abnormalities as a novel therapeutic approach in head and neck paraganglioma. Transl Oncol 14:101146 (2021). PubMed: 34118692
Li B et al. Mitochondrial-Derived Vesicles Protect Cardiomyocytes Against Hypoxic Damage. Front Cell Dev Biol 8:214 (2020). PubMed: 32426351
Liu Z et al. IFI6 depletion inhibits esophageal squamous cell carcinoma progression through reactive oxygen species accumulation via mitochondrial dysfunction and endoplasmic reticulum stress. J Exp Clin Cancer Res 39:144 (2020). PubMed: 32727517
Zhang W et al. Functional validation of a human GLUD2 variant in a murine model of Parkinson's disease. Cell Death Dis 11:897 (2020). PubMed: 33093440

View all Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Application

Flow Cytometry

Sample

Mouse Cell (Bone-marrow derived macrophages)

Permeabilization

Yes - ebio fix/perm

Specification

Bone-marrow derived macrophages

Preparation

Cell harvesting/tissue preparation method: M-CSF differentiation
Sample buffer: scraped in cold PBS

Fixation

Formaldehyde

MR. Graham Heieis

Verified customer

Submitted May 14 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (brain)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: EDTA

Permeabilization

Specification

brain

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Oct 01 2019

Application

Western blot

Sample

Mouse Tissue lysate - other (Mitochondria isolated from mouse brain)

Gel Running Conditions

Reduced Denaturing (4-12% SDS-PAGE, Laemmli buffer with 10% β-Mercaptoethanol, 10 min at +55C denaturation. Immobilon-FL transfer membranes (Millipore), overnight transfer at +4C.)

Loading amount

15 µg

Specification

Mitochondria isolated from mouse brain

Blocking step

SEA Block Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 17 2018

Application

Western blot

Loading amount

150 µg

Gel Running Conditions

Reduced Denaturing

Sample

Saccharomyces cerevisiae Cell lysate - whole cell (Yeast whole cell)

Specification

Yeast whole cell

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Dec 09 2013

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Pathway

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (14)

Customer reviews and Q&As

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Western blot abreview for Anti-SDHA antibody [EPR9043(B)]