概述

  • 产品名称

    Anti-SDHA抗体[2E3GC12FB2AE2]
    参阅全部 SDHA 一抗
  • 描述

    小鼠单克隆抗体[2E3GC12FB2AE2] to SDHA
  • 宿主

    Mouse
  • 经测试应用

    适用于: ICC, IHC-Fr, Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • 种属反应性

    与反应: Mouse, Rat, Cow, Dog, Human
  • 免疫原

    Full length native protein (purified). This information is considered to be commercially sensitive.

  • 阳性对照

    • Human heart mitochondria.
  • 常规说明

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

性能

应用

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.

应用 Ab评论 说明
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

靶标

  • 功能

    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).
  • 通路

    Carbohydrate metabolism; tricarboxylic acid cycle; fumarate from succinate (eukaryal route): step 1/1.
  • 疾病相关

    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.
  • 序列相似性

    Belongs to the FAD-dependent oxidoreductase 2 family. FRD/SDH subfamily.
  • 细胞定位

    Mitochondrion inner membrane.
  • Information by UniProt
  • 数据库链接

  • 别名

    • 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

图片

  • 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).

    Lanes 1 - 4: Merged signal (red and green). Green - ab198493 observed at 72 kDa. Red - loading control, ab181602, observed at 37 kDa.

    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).

  • 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.
  • Lane 1: Prestained Protein Ladder
    Lanes 2-3: Fly mitochondrial fraction
    Lane 4: Fly cytoplasmic fraction
    Lane 5: Robin mitochondrial fraction
    Lane 6: Robin cytoplasmic fraction
    Lane 7: Mouse (AML12) mitochondrial fraction
    Lane 8: Mouse (AML12) cytoplasmic fraction
    Lane 9: Prestained Protein Ladder

    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. 

    See Abreview

文献

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) »
See all 221 Publications for this product

客户评价及客户问答

Filter by Application

Filter by Species

Filter by Ratings

1-10 of 24 Abreviews

Application
Western blot
Sample
Caenorhabditis elegans Tissue lysate - other (Isolated Mitochondria)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
60 µg
Specification
Isolated Mitochondria
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Miss. Amber Knapp-Wilson

Verified customer

提交于 Nov 26 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA ph 9
Permeabilization
No
Specification
brain tissue
Blocking step
Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative
Formaldehyde

Herr Dr. Markus Kipp

Verified customer

提交于 Aug 22 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (SH-SY5Y)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
SH-SY5Y
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

提交于 May 24 2018

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

Abcam user community

Verified customer

提交于 Sep 12 2017

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

Abcam user community

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 Feb 05 2015

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

Abcam user community

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 Sep 03 2013

1-10 of 24 Abreviews

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

注册