Anti-ATP1A2 antibody [M7-PB-E9] (ab2871)

Overview

• Product nameAnti-ATP1A2 antibody [M7-PB-E9]See all ATP1A2 primary antibodies ...
• Description
  Mouse monoclonal [M7-PB-E9] to ATP1A2
• SpecificityDetects 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 applicationsWB, ICC/IF, Flow Cyt, IP, ELISA, IHC-Fr more details
• Species reactivity
  Reacts with: Mouse, Rat, Human
  Predicted to work with: Sheep, Chicken, Cow, Dog, Pig
• Immunogen

  Purified sheep kidney alpha sodium / potassium ATPase

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

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPBS with 0.05% sodium azide
• Concentration information loading...
• PurityAscites
• Clonality Monoclonal
• Clone numberM7-PB-E9
• IsotypeIgG1
• Research Areas
  • Metabolism
  • Types of disease
  • Cancer

  • Metabolism
  • Pathways and Processes
  • Cofactors, Vitamins / minerals
  • Vitamins / minerals

  • Signal Transduction
  • Metabolism
  • Plasma Membrane
  • ATPases

  • Signal Transduction
  • Metabolism
  • Vitamins / Minerals

Applications

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, providing the energy for active transport of various nutrients.
• Involvement in diseaseDefects in ATP1A2 are the cause of migraine familial hemiplegic type 2 (FHM2) [MIM:602481]. FHM2 is a rare, severe, autosomal dominant subtype of migraine characterized by aura and some hemiparesis.
  Defects in ATP1A2 are a cause of alternating hemiplegia of childhood (AHC) [MIM:104290]. AHC is typically distinguished from familial hemiplegic migraine by infantile onset of the symptoms and high prevalence of associated neurological deficits that become increasingly obvious with age.
• Sequence similaritiesBelongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIC subfamily.
• Cellular localizationMembrane. Cell membrane.

Target information above from: UniProt accession P50993 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 477 Human
  • Entrez Gene: 98660 Mouse
  • Entrez Gene: 24212 Rat
  • Omim: 182340 Human
  • SwissProt: P50993 Human
  • SwissProt: Q6PIE5 Mouse
  • SwissProt: P06686 Rat
  • Unigene: 34114 Human

  • Unigene: 207432 Mouse
  • Unigene: 1042 Rat
  • Unigene: 214222 Rat

  see all

• Alternative names
  AT1A2_HUMAN antibodyAtp1a2 antibodyFHM2 antibody

  KIAA0778 antibodyMHP2 antibodyNa(+)/K(+) ATPase alpha-2 subunit antibodyNa+/K+ ATPase alpha 2 subunit antibodySodium potassium ATPase antibodySodium pump subunit alpha 2 antibodySodium pump subunit alpha-2 antibodySodium/potassium transporting ATPase alpha 2 chain antibodySodium/potassium transporting ATPase subunit alpha 2 antibodySodium/potassium-transporting ATPase subunit alpha-2 antibody

  see all

Anti-ATP1A2 antibody [M7-PB-E9] images

• 

  Immunoprecipitation - Anti-ATP1A2 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) - pan alpha Sodium Potassium ATPase antibody [M7-PB-E9] (ab2871)
  This image was kindly supplied as part of the review submitted by Marko Nykanen.
• 

  Immunocytochemistry/ Immunofluorescence - Anti-ATP1A2 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.
  
  
• 

  Western blot - Anti-ATP1A2 antibody [M7-PB-E9] (ab2871)
  All lanes : Anti-ATP1A2 antibody [M7-PB-E9] (ab2871) at 1/500 dilution
  
  Lane 1 : Kidney (Human) Tissue Lysate - adult normal tissue (ab30203)
  Lane 2 : Kidney (Mouse) Tissue Lysate
  Lane 3 : Kidney (Rat) Tissue Lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (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
• 

  Flow Cytometry - Anti-ATP1A2 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 (Formalin/PFA-fixed paraffin-embedded sections)-Anti-ATP1A2 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 (Formalin/PFA-fixed paraffin-embedded sections)-Anti-ATP1A2 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 (Formalin/PFA-fixed paraffin-embedded sections)-Anti-ATP1A2 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.