Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)

Mouse monoclonal ATPB antibody [3D5]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Cow, Human, Pig, Caenorhabditis elegans, Monkey. Cited in 131 publication(s).

Reviews (28)Q&A (6)References (148)

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

    see all

  • 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

  • Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    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)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    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.
  • Flow Cytometry - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    Flow Cytometry - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    ab14730 (blue) at 1µg/ml staining ATPB in HL-60 cells and analyzed by Flow cytometry. Red histogram represents equal quantity of isotype control.
  • Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    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

  • Immunohistochemistry (Frozen sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    Immunohistochemistry (Frozen sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)This image is courtesy of an anonymous Abreview

    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

  • Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
    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:

See all 148 Publications for this product

Publishing research using ab14730? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 34 Abreviews or Q&A

Western blot abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted May 11 2015

Western blot abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Gerbil Cell lysate - whole cell (lung)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
25 µg
Specification
lung
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Jan 24 2019

Western blot abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse primary astrocytes)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Mouse primary astrocytes
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 09 2018

Immunocytochemistry abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cultured Cells (Human primary dermal fibroblasts)
Permeabilization
Yes - 0.5% TX100
Specification
Human primary dermal fibroblasts
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Paraformaldehyde
Read More

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Oct 24 2018

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
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
Read More

Mr. J. Y. Seo

Verified customer

Submitted Mar 28 2018

Immunocytochemistry abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Mar 13 2018

Immunocytochemistry abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Feb 08 2018

Western blot abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jul 27 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
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
Read More

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 23 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATPB antibody [3D5] - Mitochondrial Marker

 Excellent
Abreviews
abreview image
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
Read More

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Oct 10 2016

1-10 of 34 Abreviews or Q&A

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