Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab7671 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 5 µg/ml.
ICC/IF Use a concentration of 5 - 10 µg/ml.

We recommend Methanol fixation

WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 112 kDa. Abcam recommends using 5% BSA as the blocking agent.
Flow Cyt Use a concentration of 10 µg/ml.

See Abreview.

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

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 ions, providing the energy for active transport of various nutrients.
  • Sequence similarities
    Belongs 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 localization
    Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • A1A1 antibody
    • AT1A1 antibody
    • AT1A1_HUMAN antibody
    • ATP1A1 antibody
    • Atpa-1 antibody
    • ATPase Na+/K+ transporting alpha 1 polypeptide antibody
    • ATPase Na+/K+ transporting subunit alpha 1 antibody
    • BC010319 antibody
    • EC 3.6.3.9 antibody
    • MGC3285 antibody
    • MGC38419 antibody
    • MGC51750 antibody
    • Na K ATPase alpha A catalytic polypeptide antibody
    • Na K ATPase catalytic subunit alpha A protein antibody
    • Na(+)/K(+) ATPase 1 antibody
    • Na(+)/K(+) ATPase alpha-1 subunit antibody
    • Na+, K+ ATPase alpha subunit antibody
    • Na+/K+ ATPase alpha 1 subunit antibody
    • Na+/K+ ATPase 1 antibody
    • Na,K ATPase alpha 1 subunit antibody
    • Nkaa1b antibody
    • Sodium potassium ATPase alpha 1 polypeptide antibody
    • Sodium pump 1 antibody
    • Sodium pump subunit alpha-1 antibody
    • sodium-potassium ATPase catalytic subunit alpha-1 antibody
    • Sodium/potassium-transporting ATPase subunit alpha-1 antibody
    see all

Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] images

  • ab7671 staining alpha 1 Sodium Potassium ATPase in human formaldehyde tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% BSA for 12 hours at 4°C; antigen retrieval was by heat mediation in a buffer pH 9. Samples were incubated with primary antibody (5µg/ml in 5% BSA) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Lanes 1 - 3 : Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 1 µg/ml
    Lanes 4 - 6 : Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 2 µg/ml
    Lanes 7 - 9 : Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 5 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Brain (Mouse) Tissue Lysate (ab27253)
    Lane 3 : Brain (Rat) Tissue Lysate (ab7942)
    Lane 4 : Human brain tissue lysate - total protein (ab29466)
    Lane 5 : Brain (Mouse) Tissue Lysate (ab27253)
    Lane 6 : Brain (Rat) Tissue Lysate (ab7942)
    Lane 7 : Human brain tissue lysate - total protein (ab29466)
    Lane 8 : Brain (Mouse) Tissue Lysate (ab27253)
    Lane 9 : Brain (Rat) Tissue Lysate (ab7942)

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 112 kDa


    Exposure time : 3 minutes
  • IHC image of Ab7671 staining in Human Kidney Carcinoma 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 Ab7671, 5µg/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.
  • ab7671 staining Sodium Potassium ATPase - Plasma Membrane Marker in Human Fibrosarcoma HT1080 cells by Flow Cytometry. Cells were fixed with paraformaldehyde. The  sample was incubated with the primary antibody 10 µg/ml in PBS  for 1 hour. An Abcam PE-conjugated donkey polyclonal to mouse IgG ( ab7003), 5 µg/ml, was used as secondary antibody.

    See Abreview

  • All lanes : alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Marker at 10 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : Human kidney tissue lysate - total protein (ab30203)
    Lane 3 : Heart (Rabbit) Whole Cell Lysate - normal tissue (ab29072)
    Lane 4 : Human brain tissue lysate - total protein (ab29466)
    Lane 5 : Brain (Human) Membrane Lysate - adult normal tissue

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 112 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes
  • ab7671 staining (dog) MDCK II cells by ICC/IF.  Cells were acetone fixed and blocked with 4% serum for 10 minutes at 25°C.  The primary antibody was diluted 1/100 and incubated with the sample for 16 hours at 4°C.  Ab6785 (a FITC conjugated goat polyclonal to mouse IgG - H&L) was diluted 1/100 and employed as the secondary antibody.

    See Abreview

  • Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 1/5000 dilution + Porcine proximal tubule lysate

    Predicted band size : 112 kDa
    Western blot analysis detecting Na, K-ATP-ase (alpha) in porcine proximal tubule protein, using Ab7671. Band is at 112 kDa.
  • ICC/IF image of ab7671 stained Hek293 cells. The cells were 4% PFA fixed (10 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 (ab7671, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, HepG2 and MCF7 cells at 5µg/ml.
  • All lanes : Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 1/1000 dilution

    Lane 1 : Parotid acinar cell lysate
    Lane 2 : Parotid acinar cell lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP-conjugated Donkey anti-Mouse polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 112 kDa
    Observed band size : 112 kDa


    Exposure time : 5 minutes

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of B16 mice tumor cells labeling alpha 1 Sodium Potassium ATPase with ab7671 at 1/200 dilution. Cells were fixed in formaldehyde and permeabilized with 0.25% Triton-X100 for 10 minutes. Cells were blocked with 1% BSA for 1 hour at 20°C, followed by staining with ab7671 at 1/200 for 18 hours at 4°C in 1% BSA in PBST buffer. ab150118, a Goat Anti-Mouse IgG H&L (Alexa Fluor® 555) preadsorbed secondary antibody was used at 1/1000 dilution. DAPI was used to counterstain.

    See Abreview

References for Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671)

This product has been referenced in:
  • Pacurari M  et al. Effects of lipopolysaccharide, multiwalled carbon nantoubes, and the combination on lung alveolar epithelial cells. Environ Toxicol 32:445-455 (2017). WB, IHC ; Mouse . Read more (PubMed: 26880698) »
  • Dickens D  et al. Modulation of LAT1 (SLC7A5) transporter activity and stability by membrane cholesterol. Sci Rep 7:43580 (2017). WB ; Human . Read more (PubMed: 28272458) »

See all 131 Publications for this product

Product Wall

Thank you for your response.

Our Lab regularly test this product on whole cell lysates and heat the samples 95-100C for 10 minutes. The signal is very clear and distinctive. My personal experience is that plasmamebrane protein can be heating ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (B16 mice tumor cells)
Permeabilization
Yes - 0.25% Triton-X100 10 minutes
Specification
B16 mice tumor cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 19 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (OV90 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris-Glycin gel)
Loading amount
1e+006 cells
Specification
OV90 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Dec 06 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (N87 cell line)
Permeabilization
Yes - 0.25% Triton X-100
Specification
N87 cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 14 2016

Application
Western blot
Sample
Rat Cell lysate - whole cell (cortex)
Gel Running Conditions
Reduced Denaturing (7%)
Loading amount
10 µg
Specification
cortex
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Mar 21 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Dog Cell lysate - other (canine dermal fibroblasts-membrane fraction)
Gel Running Conditions
Reduced Denaturing (7.5%)
Loading amount
10 µg
Specification
canine dermal fibroblasts-membrane fraction
Blocking step
Block ace as blocking agent for 50 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 11 2016

Application
Western blot
Sample
Rat Cell lysate - whole cell (Lung)
Gel Running Conditions
Reduced Denaturing
Loading amount
10000 cells
Specification
Lung
Blocking step
(agent) for 30 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Oct 08 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Lung)
Specification
Lung
Blocking step
BSA as blocking agent for 1 minute(s) · Concentration: 500% · Temperature: 22°C
Fixative
Methanol
Username

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Verified customer

Submitted Jul 10 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Tissue sections (14 days old embryonic brain section, 16 micron)
Specification
14 days old embryonic brain section, 16 micron
Permeabilization
Yes - Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 23 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ph 9 steamer
Sample
Human Tissue sections (small bowel)
Specification
small bowel
Permeabilization
No
Fixative
Formaldehyde
Username

Miss. Anne Sailer

Verified customer

Submitted Dec 05 2014

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