Overview

  • Product name
    Anti-ATPB antibody [3D5] - Mitochondrial Marker
    See all ATPB primary antibodies
  • Description
    Mouse monoclonal [3D5] to ATPB - Mitochondrial Marker
  • Host species
    Mouse
  • Specificity
    Human and Bovine complex V beta subunit (ATPB).
  • Tested applications
    Suitable for: WB, IHC-Fr, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Pig, Caenorhabditis elegans, Monkey
  • Immunogen

    Whole heart mitochondria (Human).

  • General notes

    This antibody clone is manufactured by Abcam.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab14730 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 Use a concentration of 0.5 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
IHC-Fr 1/100.
ICC/IF Use a concentration of 1 - 2 µg/ml.
IP Use at an assay dependent concentration. PubMed: 21490313
Flow Cyt Use a concentration of 1 µg/ml.

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

IHC-P Use at an assay dependent concentration.

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.
  • Sequence similarities
    Belongs to the ATPase alpha/beta chains family.
  • Cellular localization
    Mitochondrion. Mitochondrion inner membrane. Peripheral membrane protein.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATP 5B antibody
    • ATP synthase H+ transporting mitochondrial F1 complex beta polypeptide antibody
    • ATP synthase subunit beta mitochondrial antibody
    • ATP synthase subunit beta, mitochondrial antibody
    • atp5b antibody
    • ATPB antibody
    • ATPB_HUMAN antibody
    • ATPMB antibody
    • ATPSB antibody
    • Epididymis secretory protein Li 271 antibody
    • HEL-S-271 antibody
    • Mitochondrial ATP synthase beta subunit antibody
    • Mitochondrial ATP Synthase Subunit Beta antibody
    • Mitochondrial ATP synthetase beta subunit antibody
    see all

Images

  • All lanes :

    Lane 1 : HeLa
    Lane 2 : HepG2
    Lane 3 : Hu liver
    Lane 4 : Ms liver

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 52 kDa



    Merged signal (red and green). Green - ab14730 observed at 52 kDa. Red - loading control, ab181602 observed at 37 kDa.

    Samples were subjected to SDS-PAGE. ab14730 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1ug/mL and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • ab14730 staining ATPB in cultured human fibroblasts. Cells were fixed, permeabilized and then labelled with ab14730 followed by an AlexaFluor® 488-conjugated Goat anti-Mouse IgG1-specific secondary antibody (2 µg/ml)
  • All lanes : Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)

    Lane 1 : isolated mitochondria from human heart at 5 µg
    Lane 2 : isolated mitochondria from bovine heart at 1 µg
    Lane 3 : isolated mitochondria from rat heart at 10 µg
    Lane 4 : isolated mitochondria from mouse heart at 10 µg

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

  • ab14730 staining ATPB in rat liver tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 10% goat serum) for 18 hours at 4°C. An Alexa Fluor® 555-conjugated goat anti-mouse polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • ab14730 (2µg/ml) staining ATPB in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic and mitochondrial staining of epithelium.
    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.
  • ab14730 (blue) at 1µg/ml staining ATPB in HL-60 cells and analyzed by Flow cytometry. Red histogram represents equal quantity of isotype control.
  • All lanes : Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730) at 0.8 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

References

This product has been referenced in:
  • Ahier A  et al. Affinity purification of cell-specific mitochondria from whole animals resolves patterns of genetic mosaicism. Nat Cell Biol N/A:N/A (2018). WB ; Caenorhabditis elegans . Read more (PubMed: 29358705) »
  • Szczesny B  et al. Mitochondrial DNA damage and subsequent activation of Z-DNA binding protein 1 links oxidative stress to inflammation in epithelial cells. Sci Rep 8:914 (2018). Read more (PubMed: 29343810) »
See all 122 Publications for this product

Customer reviews and Q&As

1-10 of 32 Abreviews or Q&A

Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (20ul of 60ul total protein extract from 4 whole fl)
Gel Running Conditions
Reduced Denaturing (4-12% BIS-TRIS PLUS)
Loading amount
1.3 cells
Specification
20ul of 60ul total protein extract from 4 whole fl
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted May 11 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
African green monkey Tissue sections (BS-c-1 cells)
Antigen retrieval step
None
Permeabilization
Yes - 0.2% Triton X
Specification
BS-c-1 cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Mr. J. Y. Seo

Verified customer

Submitted Mar 28 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Cardiomyocytes)
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 13 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Rat Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 08 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (human fibroblast cell and mitochondria fraction)
Gel Running Conditions
Reduced Denaturing
Loading amount
100 µg
Specification
human fibroblast cell and mitochondria fraction
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 27 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat primary fibroblasts)
Permeabilization
Yes - 0.5% TX100
Specification
Rat primary fibroblasts
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 23 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human primary fibroblasts)
Permeabilization
Yes - 0.5% TX100
Specification
human primary fibroblasts
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Oct 10 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Sample
Human Cell (MDA MB 231 cells)
Specification
MDA MB 231 cells
Permeabilization
Yes - 1% Triton
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 19 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Dog Cell (MDCK2 cells)
Specification
MDCK2 cells
Permeabilization
Yes - 0.5% TX100
Fixative
Paraformaldehyde

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Feb 09 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Pig Cell lysate - whole cell (porcine retinal pigment epithelium (RPE) cells)
Specification
porcine retinal pigment epithelium (RPE) cells
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jan 30 2015

1-10 of 32 Abreviews or Q&A

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