Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)

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Overview

  • Product name
    Anti-pan ATPase Alpha antibody [M7-PB-E9]
  • Description
    Mouse monoclonal [M7-PB-E9] to pan ATPase Alpha
  • Host species
    Mouse
  • Specificity
    Detects pan alpha sodium / potassium ATPase. The antibody binds all alpha subunits in sheep, dog, pig, human and chicken. It does not bind alpha 1 or 2 in rat, but does bind alpha 3.
  • Tested applications
    Suitable for: WB, ICC/IF, Inhibition Assay, ICC, Flow Cyt, IHC-P, IP, ELISA, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Chicken, Cow, Dog, Human, Pig
  • Immunogen

    Full length native protein (purified) corresponding to Sheep pan ATPase Alpha. Puified from Sheep Kidney.

  • Epitope
    This antibody recognizes an epitope between amino acid residues 646 and 652 of the sheep kidney alpha sodium / potassium ATPase.
  • Positive control
    • In western blot, this antibody gave a positive signal in human, mouse and rat kidney tissue lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab2871 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 1/500. Detects a band of approximately 102 kDa (predicted molecular weight: 114 kDa).
ICC/IF 1/20.
Inhibition Assay Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
Flow Cyt 1/100.

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

 
IHC-P Use at an assay dependent concentration.
IP 1/100 - 1/200.
ELISA Use at an assay dependent concentration.
IHC-Fr 1/100.

Target

  • Function
    This is the catalytic component of the active enzyme, which catalyzes the hydrolysis of ATP coupled with the exchange of sodium and potassium ions across the plasma membrane. This action creates the electrochemical gradient of sodium and potassium, providing the energy for active transport of various nutrients.
  • Involvement in disease
    Migraine, familial hemiplegic, 2
    Alternating hemiplegia of childhood 1
  • Sequence similarities
    Belongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIC subfamily.
  • Cellular localization
    Membrane. Cell membrane.
  • Information by UniProt
  • Database links
    • Alternative names
      • AT1A2_HUMAN antibody
      • Atp1a2 antibody
      • Na(+)/K(+) ATPase alpha-2 subunit antibody
      • Sodium pump subunit alpha-2 antibody
      • Sodium/potassium-transporting ATPase subunit alpha-2 antibody
      see all

    Images

    • Western blot - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Western blot - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      All lanes : Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871) at 1/500 dilution

      Lane 1 : Human kidney tissue lysate - total protein (ab30203)
      Lane 2 : Kidney (Mouse) Tissue Lysate
      Lane 3 : Kidney (Rat) Tissue Lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 114 kDa
      Observed band size: 102 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 28 kDa, 60 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 2 minutes
    • Immunoprecipitation - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunoprecipitation - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)

      ATP1A2 was immunoprecipitated using 0.5mg Mouse Kidney tissue extract, 5µg of Mouse monoclonal to ATP1A2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

      The antibody was incubated under agitation with Protein G beads for 10min, Mouse Kidney tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2871.

      Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

      Band: 102kDa; ATP1A2

    • Immunohistochemistry (Frozen sections) - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunohistochemistry (Frozen sections) - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      This image was kindly supplied as part of the review submitted by Marko Nykanen.
    • Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      ICC/IF image of ab2871 stained MCF7 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 (ab2871, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse 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.

    • Flow Cytometry - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Flow Cytometry - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Overlay histogram showing HEK293 cells stained with ab2871 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28711, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

      Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
    • Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha ab2871 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human testis tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha ab2871 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha ab2871 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)

      Immunocytochemistry/Immunofluorescence analysis of ATP1A2 shows staining in HeLa cells. ATP1A2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2871 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

    • Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)

      Immunocytochemistry/Immunofluorescence analysis of ATP1A2 shows staining in MCF-7 cells. ATP1A2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2871 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

    • Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)
      Immunocytochemistry/ Immunofluorescence - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)

      Immunocytochemistry/Immunofluorescence analysis of ATP1A2 shows staining in U251 cells. ATP1A2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2871 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

    References

    This product has been referenced in:
    • Ait-Omar A  et al. GLUT2 Accumulation in Enterocyte Apical and Intracellular Membranes: A Study in Morbidly Obese Human Subjects and ob/ob and High Fat-Fed Mice. Diabetes 60:2598-607 (2011). WB, IHC-Fr ; Human . Read more (PubMed: 21852673) »
    • Jiang J  et al. Localization of Na+-K+ ATPases in quasi-native cell membranes. Nano Lett 9:4489-93 (2009). Human . Read more (PubMed: 19807066) »
    See all 3 Publications for this product

    Publishing research using ab2871? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Question

    Which alpha isoform does this antibody recognize? Do we have an alpha 2 Sodium Potassium ATPase antibody?

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    Answer

    Ab2871 binds all alpha subunits in sheep, dog, pig, human and chicken. It does not bind alpha 1 or 2 in rat, but does bind alpha 3. Unfortunately we do not have a sodium potassium ATPase antibody that is specific for alpha 2. If you have any additional questions, please contact us again. Have a nice weekend!

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