Recombinant Anti-Sodium Potassium ATPase antibody [EP1845Y] - BSA and Azide free (ab167390)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1845Y] to Sodium Potassium ATPase - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human, Chinese hamster
Related conjugates and formulations
Overview
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Product name
Anti-Sodium Potassium ATPase antibody [EP1845Y] - BSA and Azide free
See all Sodium Potassium ATPase primary antibodies -
Description
Rabbit monoclonal [EP1845Y] to Sodium Potassium ATPase - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody recognizes an intracellular epitope of Sodium/potassium-transporting ATPase alpha-1 subunit.
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Chinese hamster
Predicted to work with: Tilapia -
Immunogen
Synthetic peptide corresponding to Human Sodium Potassium ATPase (N terminal).
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Positive control
- WB: HeLa, RAW 264.7, CHO, C6, MCF-7, HEK-293 and A431 whole cell lysates; Mouse brain lysate. IHC-P: Human cervical carcinoma and stomach carcinoma tissues; Mouse liver and lung tissues; Rat kidney tissue. ICC/IF: T84 cells. Flow Cyt (intra): HeLa cells.
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General notes
ab167390 is the carrier-free version of ab76020.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1845Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- HRP Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab185065)
- Alexa Fluor® 488 Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Marker (ab197713)
- Biotin Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab198366)
- Alexa Fluor® 647 Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Marker (ab198367)
- PE Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab209299)
- Alexa Fluor® 405 Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab210143)
- Alexa Fluor® 555 Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab274883)
- APC Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab314288)
- Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020)
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab167390 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
Please refer to the original abID, ab76020, for information on recommended dilutions. |
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ICC/IF |
Use at an assay dependent concentration.
Please refer to the original abID, ab76020, for information on recommended dilutions. |
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Please refer to the original abID, ab76020, for information on recommended dilutions. |
WB |
Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 113 kDa).
Please refer to the original abID, ab76020, for information on recommended dilutions. |
Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. Please refer to the original abID, ab76020, for information on recommended dilutions. |
ICC/IF
Use at an assay dependent concentration. Please refer to the original abID, ab76020, for information on recommended dilutions. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Please refer to the original abID, ab76020, for information on recommended dilutions. |
WB
Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 113 kDa). Please refer to the original abID, ab76020, for information on recommended dilutions. |
Target
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 476 Human
- Entrez Gene: 480 Human
- Entrez Gene: 481 Human
- Entrez Gene: 11928 Mouse
- Entrez Gene: 11931 Mouse
- Entrez Gene: 27222 Mouse
- Entrez Gene: 24211 Rat
- Entrez Gene: 25650 Rat
see all -
Alternative names
- ATPase Na+/K+ transporting alpha antibody
- adenosinetriphosphatase antibody
- AT1A1_HUMAN antibody
see all
Images
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All lanes : Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/100000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared from RIPA lysis method
Lane 2 : HeLa whole cell lysate prepared from 1% SDS HOT lysis method
Lane 3 : HeLa whole cell lysate prepared from RIPA lysis method
Lane 4 : HeLa whole cell lysate prepared from 1%SDS HOT lysis method
Lane 5 : Raw264.7 (Mouse abelson murine leukemia virus-induced tumor) whole cell lysate prepared from RIPA lysis method
Lanes 6 & 8 : Raw264.7 whole cell lysate prepared from 1%SDS HOT lysis method
Lane 7 : Raw264.7 whole cell lysate prepared from RIPA lysis method
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Predicted band size: 113 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThis data was produced using ab76020, the same antibody clone in a different formulation
Blocking/Diluting buffer and concentration 5% NFDM/TBST
We suggest not to boil the sample after lysis.
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T84 cells cultured on 8-well chamber slides, were washed once with ice-cold PBS, then fixed with 4% paraformaldehyde for 30 min at 4°C. After fixation, cells were permeabilized with 0.5% Triton X-100 for 5 min at room temperature and washed with PBS three times. Following blocking with 2% FCS in PBS for 1 hour at room temperature, primary antibody staining was performed at 4°C overnight at 1/200 dilution. Cells were then incubated with protein fractions B12 and C5 at 5x dilutions in fresh media for 1 hour at 37°C. Cells were then fixed, permeabilized and co-stained with fiber and sodium potassium ATPase. The nuclei were stained with DAPI using Vectachield mounting medium. Cells were visualized using Zeiss confocal microscopy LSM700.
Fiber molecules were found to be predominantly intracellularly following B12 treatment.
For full image see PubMed: 25723153.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
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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 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
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Anti-Sodium Potassium ATPase antibody [EP1845Y] - BSA and Azide free (ab167390) + MCF-7 (human breast carcinoma) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Predicted band size: 113 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 8 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
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All lanes : Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/100000 dilution
Lane 1 : CHO (Chinese hamster ovary cell line) cell lysate
Lane 2 : C6 (Rat glial tumor cell line) cell lysate
Lane 3 : Mouse brain
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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?This data was produced using ab76020, the same antibody clone in a different formulation.
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
We suggest not to boil the sample after lysis.
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Clone EP1845Y (ab167390) has been successfully conjugated by Abcam. This image was generated using Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Marker (Alexa Fluor® 488). Please refer to ab197713 for protocol details.
ab197713 staining Sodium Potassium ATPase in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab197713 at a 1/50 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Clone EP1845Y (ab167390) has been successfully conjugated by Abcam. This image was generated using Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Marker (Alexa Fluor® 647). Please refer to ab198367 for protocol details.
ab198367 staining Sodium Potassium ATPase in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab198367 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
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All lanes : Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/100000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 4 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution
Predicted band size: 113 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutesThis data was produced using ab76020, the same antibody clone in a different formulation.
Blocking and diluting buffer: 5% NFDM/TBST.
We suggest not to boil the sample after lysis.
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Clone EP1845Y (ab167390) has been successfully conjugated by Abcam. This image was generated using Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (PE). Please refer to ab209299 for protocol details.
Overlay histogram showing HeLa cells stained with ab209299 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab209299, 1/2500 dilution) for 30 min at 22°C.
Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 50mW Yellow-Green laser (561nm) and 586/15 bandpass filter.
This antibody gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76020).
Protocols
Datasheets and documents
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Datasheet download
References (6)
ab167390 has been referenced in 6 publications.
- Hargest V et al. Astrovirus-induced epithelial-mesenchymal transition via activated TGF-β increases viral replication. PLoS Pathog 18:e1009716 (2022). PubMed: 35452499
- Izem L et al. Plasminogen-induced foam cell formation by macrophages occurs through a histone 2B (H2B)-PAR1 pathway and requires integrity of clathrin-coated pits. J Thromb Haemost 19:941-953 (2021). PubMed: 33492784
- Ferrian S et al. Multiplexed imaging reveals an IFN-γ-driven inflammatory state in nivolumab-associated gastritis. Cell Rep Med 2:100419 (2021). PubMed: 34755133
- Cheung AM et al. Quantitative single-cell analysis of immunofluorescence protein multiplex images illustrates biomarker spatial heterogeneity within breast cancer subtypes. Breast Cancer Res 23:114 (2021). PubMed: 34922607
- Wang J et al. DHHC4 and DHHC5 Facilitate Fatty Acid Uptake by Palmitoylating and Targeting CD36 to the Plasma Membrane. Cell Rep 26:209-221.e5 (2019). PubMed: 30605677
- Huang C et al. Acetylation within the N- and C-Terminal Domains of Src Regulates Distinct Roles of STAT3-Mediated Tumorigenesis. Cancer Res 78:2825-2838 (2018). PubMed: 29531159