Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab76020 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
ICC/IF 1/500.
WB 1/100000. Predicted molecular weight: 113 kDa.

For unpurified, use 1/20000.

Flow Cyt 1/20 - 1/100.

Follow an intracellular staining protocol.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Target

  • FunctionThis 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 ions, providing the energy for active transport of various nutrients.
  • Sequence similaritiesBelongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIC subfamily.
  • Post-translational
    modifications
    Phosphorylation on Tyr-10 modulates pumping activity.
  • Cellular localizationCell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATPase Na+/K+ transporting alpha antibody
    • adenosinetriphosphatase antibody
    • AT1A1_HUMAN antibody
    • ATP1A1 antibody
    • ATP1A4 antibody
    • ATP1AL2 antibody
    • ATP1B antibody
    • ATP1B1 antibody
    • ATPase Na+/K+ transporting alpha 1 polypeptide antibody
    • ATPase Na+/K+ transporting alpha 4 polypeptide antibody
    • ATPase Na+/K+ transporting beta 1 polypeptide antibody
    • ATPase, Na+/K+ transporting, alpha polypeptide-like 2 antibody
    • ATPase, Na+/K+ transporting, beta 1 polypeptide antibody
    • Beta 1-subunit of Na(+),K(+)-ATPase antibody
    • Na(+)/K(+) ATPase alpha-1 subunit antibody
    • Na(+)/K(+) ATPase alpha-4 subunit antibody
    • Na+, K+ ATPase alpha subunit antibody
    • Na+/K+ ATPase 1 antibody
    • Na+/K+ ATPase 4 antibody
    • Na+/K+ ATPase, alpha-D polypeptide antibody
    • Na, K-ATPase beta-1 polypeptide antibody
    • Na, K-ATPase, alpha-A catalytic polypeptide antibody
    • Na,K-ATPase catalytic subunit alpha-A protein antibody
    • Na,K-ATPase subunit alpha-C antibody
    • polypeptide-like 2 antibody
    • Sodium pump 1 antibody
    • sodium pump 4 antibody
    • Sodium pump subunit alpha-1 antibody
    • sodium pump subunit alpha-4 antibody
    • sodium pump subunit beta-1 antibody
    • sodium-potassium ATPase catalytic subunit alpha-1 antibody
    • sodium-potassium ATPase catalytic subunit alpha-4 antibody
    • sodium-potassium ATPase subunit beta 1 (non-catalytic) antibody
    • sodium-potassium ATPase, alpha 4 polypeptide antibody
    • sodium-potassium-ATPase, alpha 1 polypeptide antibody
    • Sodium/potassium transporting ATPase alpha 1 chain antibody
    • Sodium/potassium transporting ATPase subunit beta 1 antibody
    • sodium/potassium-dependent ATPase beta-1 subunit antibody
    • sodium/potassium-transporting ATPase alpha-4 chain antibody
    • sodium/potassium-transporting ATPase beta-1 chain antibody
    • Sodium/potassium-transporting ATPase subunit alpha-1 antibody
    • sodium/potassium-transporting ATPase subunit alpha-4 antibody
    see all

Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control images

  • ab76020 staining Sodium Potassium ATPase in lung epithelia (top) and bronchiolar epithelia (bottom) from Mouse lung tissue sections by Immunohistochemistry ((IHC) - paraffin-embedded sections). Sections were deparaffined at 60°C and rehydrated by successive incubations in 100% xylol, 100% ethanol and 96% ethanol. Samples were then permeabilized with 0.25% Triton X-100 in PBS for 15 minutes and blocked with 10% bovine serum albumin (BSA) and 0.25% Triton X-100 in PBS for 30 minutes. Samples were incubated with primary antibody (1/100 in 0.25% Triton X-100 and 10% BSA in PBS) for 1 hour 30 minutes at room temperature. An Alexa Fluor®488-conjugated Goat anti-mouse antibody was used as the secondary antibody.

  • All lanes : Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/100000 dilution

    Lane 1 : CHO cell lysate
    Lane 2 : C6 cell lysate
    Lane 3 : Mouse brain

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 113 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) cells labelling Sodium Potassium ATPase with purified ab76020 at 1/500. Cells were fixed with 100% methanol. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

  • Overlay histogram showing HeLa cells fixed in 80% methanol and stained with purified ab76020 at a dilution of 1 in 100 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control (black line) and the blue line shows cells incubated without primary or secondary antibody.

  • All lanes : Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/100000 dilution (purified)

    Lane 1 : MCF7 cell lysate
    Lane 2 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 113 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded mouse liver with purified ab76020 at a working dilution of 1 in 100. The secondary antibody used is a HRP conjugated goat anti-rabbit IgG (H+L), ab97051, at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded rat kidney with purified ab76020 at a working dilution of 1 in 100. The secondary antibody used is a HRP conjugated goat anti-rabbit IgG (H+L), ab97051, at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab76020 at a working dilution of 1 in 100. The secondary antibody used is a HRP conjugated goat anti-rabbit IgG (H+L), ab97051, at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/20000 dilution (unpurified) + Hela cell lysate at 10 µg

    Secondary
    HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size : 113 kDa
  • Immunohistochemical staining of Sodium Potassium ATPase in paraffin embedded human stomach carcinoma tissue with unpurified ab76020, at a 1/100 dilution.

  • Overlay histogram showing HeLa cells stained with unpurified ab76020 (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 (ab76020, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/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.

References for Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020)

This product has been referenced in:
  • Marshall DJ  et al. Extracellular Antibody Drug Conjugates Exploiting the Proximity of Two Proteins. Mol Ther N/A:N/A (2016). WB . Read more (PubMed: 27434591) »
  • Zhao LL  et al. Insulin-independent GLUT4 translocation in proliferative vascular smooth muscle cells involves SM22a. J Mol Med (Berl) N/A:N/A (2016). Read more (PubMed: 27631639) »

See all 17 Publications for this product

Product Wall

Application Western blot
Sample Ferret Tissue lysate - whole (Whole Brain tissue (Membrane proteins extracted an)
Gel Running Conditions Reduced Denaturing (3-8% Tris-acetate gel, NuPage)
Loading amount 40 µg
Specification Whole Brain tissue (Membrane proteins extracted an
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Francis Djankpa

Verified customer

Submitted Jul 11 2016

Application Western blot
Sample Mouse Cell lysate - other (membrane fraction bEnd.3)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification membrane fraction bEnd.3
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Sep 15 2015

Thank you for your inquiry. I can confirm that the concetration of the batch currently in stock is xxxx mg/mL. I also would like to comment on the seemingly low concentration: The IgG produced by rabbits have greater affinity and greater sensitivity th...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"