Recombinant Anti-Sodium Potassium ATPase antibody [EP1845Y] - Rat IgG2a (Chimeric) (ab283345)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rat monoclonal [EP1845Y] to Sodium Potassium ATPase - Rat IgG2a
- Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Sodium Potassium ATPase antibody [EP1845Y] - Rat IgG2a (Chimeric)
See all Sodium Potassium ATPase primary antibodies -
Description
Rat monoclonal [EP1845Y] to Sodium Potassium ATPase - Rat IgG2a -
Host species
Rat -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
The details of the immunogen for this antibody are not available.
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Positive control
- WB: HeLa, HeLa prepared from 1% SDS HOT lysis method, HeLa prepared from RIPA lysis method, RAW264.7 and C6 whole lysates. IHC-P: Human colon, Human hepatocellular carcinoma, Mouse kidney, Mouse stomach and Rat kidney, Rat stomach tissues. ICC: HeLa cells. Flow Cyt: HeLa cell.
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General notes
This rat monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (ab76020). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.
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
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Ion Exchange Chromatography -
Clonality
Monoclonal -
Clone number
EP1845Y -
Isotype
IgG2a -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283345 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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WB |
1/1000. Predicted molecular weight: 113 kDa.
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/100.
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ICC/IF |
1/50.
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Notes |
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WB
1/1000. Predicted molecular weight: 113 kDa. |
IHC-P
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/100. |
ICC/IF
1/50. |
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] - Rat IgG2a (Chimeric) (ab283345) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate prepared from RIPA lysis method
Lanes 2 & 4 : HeLa whole cell lysate prepared from 1% SDS HOT lysis method
Lane 3 : HeLa whole cell lysate prepared from RIPA lysis method
Lane 5 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 6 : RAW264.7 whole cell lysate
Lane 7 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 8 : C6 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG (H+L), HRP) (ab205720) at 1/5000 dilution
Predicted band size: 113 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Samples of Lanes 3, 4, 6, 8 are non-boiled as boiling may cause protein aggregates.
Exposure time: 1 second
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Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Sodium Potassium ATPase with ab283345 at 1/5000 (0.222 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on human colon. The section was incubated with ab283345 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling Sodium Potassium ATPase with ab283345 at 1/5000 (0.222 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on human hepatocellular carcinoma. The section was incubated with ab283345 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Sodium Potassium ATPase with ab283345 at 1/5000 (0.222 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Mainly membranous staining on mouse kidney. The section was incubated with ab283345 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling Sodium Potassium ATPase with ab283345 at 1/5000 (0.222 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on mouse stomach. The section was incubated with ab283345 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Sodium Potassium ATPase with ab283345 at 1/5000 (0.222 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Mainly membranous staining on rat kidney. The section was incubated with ab283345 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Sodium Potassium ATPase with ab283345 at 1/5000 (0.222 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on rat stomach. The section was incubated with ab283345 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Sodium Potassium ATPase with ab283345 at 1/50 (22.18 ug/ml) dilution, followed by ab150161 Goat F(ab')2 Anti-Rat IgG Fc (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in HeLa cell line. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150161 Goat F(ab')2 Anti-Rat IgG Fc (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Sodium Potassium ATPase with ab283345 at 1/100 dilution (1ug) (Red) compared with a Rat monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor® 488, ab150161) at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab283345 has not yet been referenced specifically in any publications.