Overview

  • Product name
    Anti-ATP5A antibody [EPR13030(B)]
    See all ATP5A primary antibodies
  • Description
    Rabbit monoclonal [EPR13030(B)] to ATP5A
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ATP5A aa 200-300 (Cysteine residue). The exact sequence is proprietary. Isoform 1
    Database link: P25705

  • Positive control
    • HepG2, HeLa, fetal liver and fetal lung lysates; Human liver and fetal heart tissues; HeLa and MCF7 cells.
  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab176569 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
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/100 - 1/250.
IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/50 - 1/100.

The use of an HRP/AP polymerized secondary antibody is recommended.

WB 1/1000 - 1/10000. Predicted molecular weight: 60 kDa.

Target

  • Function
    Mitochondrial membrane ATP synthase (F(1)F(0) ATP synthase or Complex V) produces ATP from ADP in the presence of a proton gradient across the membrane which is generated by electron transport complexes of the respiratory chain. F-type ATPases consist of two structural domains, F(1) - containing the extramembraneous catalytic core, and F(0) - containing the membrane proton channel, linked together by a central stalk and a peripheral stalk. During catalysis, ATP synthesis in the catalytic domain of F(1) is coupled via a rotary mechanism of the central stalk subunits to proton translocation. Subunits alpha and beta form the catalytic core in F(1). Rotation of the central stalk against the surrounding alpha(3)beta(3) subunits leads to hydrolysis of ATP in three separate catalytic sites on the beta subunits. Subunit alpha does not bear the catalytic high-affinity ATP-binding sites.
  • Tissue specificity
    Fetal lung, heart, liver, gut and kidney. Expressed at higher levels in the fetal brain, retina and spinal cord.
  • Sequence similarities
    Belongs to the ATPase alpha/beta chains family.
  • Post-translational
    modifications
    The N-terminus is blocked.
  • Cellular localization
    Mitochondrion inner membrane. Peripheral membrane protein.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATP synthase alpha chain, mitochondrial antibody
    • ATP synthase subunit alpha antibody
    • ATP synthase subunit alpha mitochondrial antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1, cardiac muscle antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, 1 antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, isoform 1, cardiac muscle antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, isoform 2, non-cardiac muscle-like 2 antibody
    • ATP sythase (F1 ATPase) alpha subunit antibody
    • ATP5A antibody
    • Atp5a1 antibody
    • ATP5AL2 antibody
    • ATPA_HUMAN antibody
    • ATPM antibody
    • Epididymis secretory sperm binding protein Li 123m antibody
    • hATP1 antibody
    • HEL-S-123m antibody
    • MC5DN4 antibody
    • mitochondrial antibody
    • Mitochondrial ATP synthetase antibody
    • Mitochondrial ATP synthetase oligomycin resistant antibody
    • Modifier of Min 2 mouse homolog antibody
    • Modifier of Min 2, mouse, homolog of antibody
    • MOM2 antibody
    • OMR antibody
    • ORM antibody
    • OTTHUMP00000163475 antibody
    see all

Images

  • All lanes : Anti-ATP5A antibody [EPR13030(B)] (ab176569) at 0.01 µg/ml

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 60 kDa



    Blocking and diluting buffer: 5% NFDM/TBST
  • Ab176569 (purified) staining ATP5A in HeLa (human cervix adenocarcinoma epithelial cell) by Immunocytochemistry/Immunofluorescence (ICC/IF). Cells were fixed with 4% paraformaldehyde and permeabilized n 0.1% TritonX-100. Samples were incubated with primary antibody at 1/500 dilution (4.2µg/ml).  An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1/1000 dilution (2µg/ml). DAPI was used as a nuclear counterstain. Confocal image showing cytoplasmic staining in HeLa cells.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling ATP5A with Purified ab176569 at 1:500 dilution (0.21 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling ATP5A with Purified ab176569 at 1:500 dilution (0.21 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling ATP5A with Purified ab176569 at 1:500 dilution (0.21 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylin was used as a counterstain.
  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ATP5A with purified ab176569 at 1:60 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • All lanes : Anti-ATP5A antibody [EPR13030(B)] (ab176569) at 1/1000 dilution (unpurified)

    Lane 1 : HepG2 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Human fetal liver lysate
    Lane 4 : Human fetal lung lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Developed using the ECL technique.

    Predicted band size: 60 kDa

  • Immunofluorescence analysis of MCF7 cells labeling ATP5A using ab176569 (unpurified) at a 1/100 dilution (green). DAPI nuclear staining (blue).

  • Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling ATP5A using ab176569 (unpurified) at a 1/50 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human fetal heart tissue labeling ATP5A using ab176569 (unpurified) at a 1/50 dilution.

  • Flow cytometric analysis of permeabilized HeLa cells labeling ATP5A using ab176569 (unpurified) at a 1/10 dilution (red) or a rabbit IgG negative control (green).

References

This product has been referenced in:
  • Song W  et al. Organic cation transporter 3 (Oct3) is a distinct catecholamines clearance route in adipocytes mediating the beiging of white adipose tissue. PLoS Biol 17:e2006571 (2019). Read more (PubMed: 30653498) »
  • Zhu M  et al. Mitochondria Released by Apoptotic Cell Death Initiate Innate Immune Responses. Immunohorizons 2:384-397 (2018). Read more (PubMed: 30847435) »
See all 6 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (A549)
Permeabilization
Yes - 0.1% Triton X-100
Specification
A549
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Nov 06 2017

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse cells lines (3T3 fibroblasts, C2C12 myoblast)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
10 µg
Specification
Mouse cells lines (3T3 fibroblasts, C2C12 myoblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 21 2017

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