Overview

  • Product name

  • Description

    Rabbit polyclonal to ATPB
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Saccharomyces cerevisiae
    Predicted to work with: Rabbit, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Chinese hamster
  • Immunogen

    Synthetic peptide corresponding to Human ATPB aa 150-250 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab140747)

  • Positive control

    • This antibody gave a positive signal in Mouse Liver tissue lysate as well as the following whole cell lysates: HeLa; HepG2; MEF1; NIH3T3; Raw264.7; PC12. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal heart muscle. This antibody gave a positive result when used in the following methanol fixed cell lines: HepG2.

Properties

Applications

Our Abpromise guarantee covers the use of ab128743 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
IHC-P Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
ICC/IF Use a concentration of 5 µg/ml.

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 : Anti-ATPB antibody (ab128743) at 1 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 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
    Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 7 : Liver (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 56 kDa
    Observed band size: 56 kDa


    Exposure time: 10 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab128743 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
  • ab128743 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab128743 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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) at a concentration of 1.43µM.

  • IHC image of ATPB staining in human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab128743, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-ATPB antibody (ab128743) at 1/5000 dilution

    Lane 1 : Saccharomyces cerevisiae strain BY4741 whole cell lysate
    Lane 2 : Saccharomyces cerevisiae strain BY4741 delta ATP2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG monoclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 56 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 130 kDa (possible non-specific binding)


    Exposure time: 5 minutes

    See Abreview

References

This product has been referenced in:

  • Du C  et al. Isobaric tags for relative and absolute quantitation-based proteomics reveals potential novel biomarkers for the early diagnosis of acute myocardial infarction within 3 h. Int J Mol Med 43:1991-2004 (2019). Read more (PubMed: 30896787) »
  • Pereira C  et al. Sit4p-mediated dephosphorylation of Atp2p regulates ATP synthase activity and mitochondrial function. Biochim Biophys Acta 1859:591-601 (2018). Saccharomyces cerevisiae . Read more (PubMed: 29719209) »
See all 4 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (12.5% Acrylamide)
Sample
Pig Tissue lysate - other (Muscle Sarcoplasm)
Specification
Muscle Sarcoplasm
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Shannon Cruzen

Verified customer

Submitted Sep 30 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12,5%)
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (strain BY4741)
Specification
strain BY4741
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 11 2014

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