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

Overview

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

  • FunctionMitochondrial 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 similaritiesBelongs to the ATPase alpha/beta chains family.
  • Cellular localizationMitochondrion. 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

Anti-ATPB antibody [3D5] - Mitochondrial Marker images

  • 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.
  • ab14730 staining ATPB in the mouse MEF cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with PFA, permeabilized with 1% Triton X-100 and blocked with 5% goat serum for 1 hour at 21°C. Samples were incubated with primary antibody (1/200 in 5% goat serum) for 2 hours at 21°C. An  Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody.

    See Abreview

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

This product has been referenced in:
  • Bakshi MV  et al. In-Utero Low-Dose Irradiation Leads to Persistent Alterations in the Mouse Heart Proteome. PLoS One 11:e0156952 (2016). WB ; Mouse . Read more (PubMed: 27276052) »
  • Tome ME  et al. Identification of P-glycoprotein co-fractionating proteins and specific binding partners in rat brain microvessels. J Neurochem 134:200-10 (2015). Read more (PubMed: 25832806) »

See all 63 Publications for this product

Product Wall

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
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Submitted May 11 2015

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
Username

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
Username

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
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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
Username

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
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jan 30 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (15%)
Sample Human Cell lysate - whole cell (human primary fibroblasts)
Specification human primary fibroblasts
Blocking step Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jan 29 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (15%)
Sample Dog Cell lysate - whole cell (MDCK2 cells)
Specification MDCK2 cells
Blocking step Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jan 21 2015

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

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jan 13 2015

Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Human Cell lysate - other (mitochondrial extract from HEK293 cells)
Specification mitochondrial extract from HEK293 cells
Total protein in input 200 µg
Username

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Verified customer

Submitted Nov 17 2014

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