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Anti-Alpha 2 Na+/K+ ATPase Antibody

ATP1A2, Sodium/potassium ATPase catalytic subunit alpha-2, Sodium pump subunit alpha-2

Cat #: ANP-002
Alternative Name ATP1A2, Sodium/potassium ATPase catalytic subunit alpha-2, Sodium pump subunit alpha-2
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)REYSPAATTAENG, corresponding to amino acid residues 7-19 of rat ATP1A2 (Accession P06686). Intracellular, N-terminus.
Accession (Uniprot) Number P06686
Gene ID 24212
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse, human, dog, pig – identical.
RRID AB_2756680.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ih, wb
May also work in: ic*, ifc*, ip*
Western blot
  • Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
    Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
    1,2. Anti-Alpha 2 Na+/K+ ATPase Antibody (#ANP-002), (1:200).
    3,4. Anti-Alpha 2 Na+/K+ ATPase Antibody, preincubated with Alpha 2 Na+/K+ ATPase Blocking Peptide (#BLP-NP002).
  • Western blot analysis of human U-87 MG glioblastoma cell lysate:
    Western blot analysis of human U-87 MG glioblastoma cell lysate:
    1. Anti-Alpha 2 Na+/K+ ATPase Antibody (#ANP-002), (1:200).
    2. Anti- Alpha 2 Na+/K+ ATPase Antibody, preincubated with Alpha 2 Na+/K+ ATPase Blocking Peptide (#BLP-NP002).
Immunohistochemistry
  • Expression of Na+/K+ ATPase α2 (ATP1A2) in mouse hippocampus and cerebellum
    Expression of Na+/K+ ATPase α2 (ATP1A2) in mouse hippocampus and cerebellum
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Alpha 2 Na+/K+ ATPase Antibody (#ANP-002), (1:300), followed by goat-anti-rabbit-AlexaFluor-488 secondary antibody. A. ATP1A2 staining (green) in mouse hippocampal CA3 region is labeled in astrocytic processes (arrows surrounding pyramidal cells, P) and diffused staining is observed in the CA3 stratum radiatum. Note absence of staining in the mossy fiber terminal region (MF). B. Staining in mouse cerebellum shows labeling of the basket shape (arrows) under Purkinje cells (P) and diffused staining in the molecular layer (MOL). Cell nuclei are stained with DAPI (blue).
References
  1. Jorgensen, P.L. et al. (2003) Annu. Rev. Physiol. 65, 817.
  2. Kaplan, J.H. (2002) Annu. Rev. Biochem. 71, 511.
  3. Friedrich, T. et al. (2016) Front. Physiol. 7, 239.
Scientific background

The Na+/K+ pumps are members of the P-type family of active cation transport proteins. They consume ATP to actively transport Na+ out of and K+ into the cell. Na+ and K+ gradients are required for maintaining membrane potential, cell volume and secondary transport of other solutes thus making these pumps essential for all mammalian cells.
 
The minimum functional unit of the Na+/K+ pump consists of a heterodimer of α- and β- subunits1. The α subunit (also known as ATP1A) has a molecular mass of about 100 kDa and four distinct isoforms have been identified so far. It has ten transmembrane segments, short extracellular loops and larger cytoplasmic regions. There is a large central loop between M4 and M5 composed of 430 amino acid residues and a long N-terminal tail of about 90 amino acid residues. The nucleotide-binding domain (N domain) and phosphorylation domain (P domain) of the pump are formed by the M4/M5 loop while the actuator domain (A domain) is formed by the amino-terminal tail and the M2/M3 loop. The major isoform, α1, is found in most tissues and is the major form in the kidney and in most epithelia cells. The β-subunit is essential for the delivery and insertion of the α-subunit into the cell’s membrane, whilst β-subunit delivery to the membrane is not dependent on α-subunit expression2.
 
ATP1A2 mutations that cause diverse functional abnormalities of the Na+/K+ pump have been implicated in the pathogenesis of familial hemiplegic migraine3.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 24/01/2021

Anti-Alpha 2 Na+/K+ ATPase Antibody (#ANP-002) is a highly specific antibody directed against an epitope of the rat ATP1A2. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize ATP1A2 from human, rat, and mouse samples.

For research purposes only, not for human use
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