Anti-Beta 2 Na+/K+ ATPase (ext

Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular)

ATPase Na⁺/K⁺ Transporting Subunit Beta 2, Adhesion Molecule On Glia, AMOG, ATP1B2
Cat #: ANP-012
Sizes: 25 µl, 50 µl, 0.2 ml
Source: Rabbit
Type: Polyclonal
Applications: IH, WB
May also work in: IC, IFC, IP, LCI
Reactivity: H, M, R

Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular)Application key:

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

Species reactivity key:

H- Human, M- Mouse, R- Rat
CLICK HERE TO RECEIVE A 25 µl FREE TRIAL SAMPLE!

Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular) (#ANP-012) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot analysis. The antibody recognizes an extracellular epitope, and could potentially be used for detecting the protein in living cells. It has been designed to recognize Beta 2 Na⁺/K⁺ ATPase from rat, mouse, and human samples.

• Applications
• Specifications
• Scientific Background
• Related Products

Western blot

Western blot analysis of rat brain lysate:
1. Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular) antibody (#ANP-012), (1:400).
2. Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular) antibody, preincubated with the control peptide antigen.

Western blot analysis of mouse brain lysate:
1. Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular) antibody (#ANP-012), (1:200).
2. Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular) antibody, preincubated with the control peptide antigen.
Immunohistochemistry

Expression of Beta 2 Na⁺/K⁺ ATPase in mouse hippocampus
Immunohistochemical staining of perfusion-fixed frozen mouse brain sections with Anti-Beta 2 Na⁺/K⁺ ATPase (extracellular) antibody (#ANP-012), (1:300), followed by goat anti-rabbit-AlexaFluor-488. Beta 2 Na⁺/K⁺ ATPase staining (green) in the hippocampal CA1 region, is detected in glial cell profiles (horizontal arrows) in the stratum radiatum (SR) and pyramidal (Pyr) layer. Cell nuclei are stained with DAPI (blue).
Immunogen
Peptide (C)DTESWDQHVQKLNK, corresponding to amino acid residues 99-112 of rat Beta 2 Na⁺/K⁺ ATPase (Accession P13638). Extracellular, C-terminus.

HomologyHuman – identical; mouse 13/14 amino acid residues identical.
PurityAffinity purified on immobilized antigen.
FormulationLyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.
Standard quality control of each lotWestern blot analysis.
Peptide confirmationConfirmed by amino acid analysis and mass spectrometry.
Storage before reconstitutionThe antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution0.8 mg/ml.
Storage after reconstitutionThe 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).
Control antigen storage before reconstitutionLyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
Control antigen reconstitution100 µl double distilled water (DDW).
Control antigen storage after reconstitution-20ºC.
Preadsorption Control1 µg peptide per 1 µg antibody.
Scientific background

P-type ATPases are a large family of molecular pumps that exploit a phosphorylated enzyme intermediate in a two-step mechanism of ATP hydrolysis, cycling through states which are associated with ion transport or ion counter-transport. The Na⁺/K⁺-ATPase, a member of this family, is almost exclusively found in animals, although close homologues have been reported in certain archaea, algae and oomycetes.

The Na⁺/K⁺-ATPase is comprised of a nucleotide-binding (N) and phosphorylation (P) domain, a transmembrane core (M1–M6) and a large carbozk-terminal M7–M10 segment. Na⁺/K⁺-ATPase undergoes large conformational changes as part of its functional cycle giving rise to two distinct enzymatic states: E1, which is a high-affinity state for the primary transported ion- Na⁺ and E2, which is the low-affinity state for the Na⁺ ion. The two states arise from the autocatalysed formation and breakdown of a phosphoenzyme intermediate, coupled to the binding, occlusion, translocation and release of ions. The phosphorylation site is the Asp residue of a conserved DKTGT motif. The core of the membrane transport domain encompasses transmembrane helices M1–M6, which hold the main ion-binding sites and are necessary for cytoplasmic and extracellular ion transport. A terminal R domain extension, which serves as a regulatory unit, can be found in the C-terminal of the protein. This unit is auto-inhibitory and is predicted to restrict transmembrane helix movements and/or access of ions to the membrane transport core¹.  

Na⁺/K⁺-ATPase has been implicated in the pathogenesis of Alzheimer’s Disease. A deficiency in several Na⁺/K⁺-ATPase isoform genes induce learning and memory deficits, and the α isoform is altered in Alzheimer’s Disease².

References

1. Morth, J.P. et al. (2011) Nat. Rev. Mol. Cell Biol. 12, 60.
2. Zhang, L.N. et al. (2012) Fundam. Clin. Pharmacol. 27, 96.