Na,K-ATPase Antibody

3010:

Western Blotting

Na,K-ATPase α Antibody detects endogenous levels of total Na,K-ATPase α1 protein. Based on sequence homology, the antibody is likely to cross-react with α2 and α3 isoforms. A doublet may form if samples are boiled.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Na,K-ATPase α1 subunit. Antibodies are purified using peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from various cell types using Na,K-ATPase α Antibody.

The Na,K-ATPase is an integral membrane heterodimer belonging to the P-type ATPase family. This ion channel uses the energy derived from ATP hydrolysis to maintain membrane potential by driving sodium export and potassium import across the plasma membrane against their electrochemical gradients. It is composed of a catalytic α subunit and a β subunit (reviewed in 1). Several phosphorylation sites have been identified for the α1 subunit. Tyr10 is phosphorylated by an as yet undetermined kinase (2), Ser16 and Ser23 are phosphorylated by PKC, and Ser943 is phosphorylated by PKA (3-5). All of these sites have been implicated in the regulation of enzyme activity in response to hormones and neurotransmitters, altering trafficking and kinetic properties of Na,K-ATPase. Altered phosphorylation in response to angiotensin II stimulates activity in the rat proximal tubule (6). Na,K-ATPase is also involved in other signal transduction pathways. Insulin regulates its localization in differentiated primary human skeletal muscle cells, and this regulation is dependent on ERK1/2 phosphorylation of the α subunit (7). Na,K-ATPase and Src form a signaling receptor complex that affects regulation of Src kinase activity and, subsequently, its downstream effectors (8,9).

1. Therien, A.G. and Blostein, R. (2000) Am. J. Physiol. Cell Physiol. 279, C541-566.
2. Féraille, E. et al. (1999) Mol. Biol. Cell 10, 2847-2859.
3. Fisone, G. et al. (1994) J. Biol. Chem. 269, 9368-9373.
4. Feschenko, M.S. and Sweadner, K.J. (1995) J. Biol. Chem. 270, 14072-14077.
5. Beguin, P. et al. (1994) J. Biol. Chem. 269, 24437-24445.
6. Yingst, D.R. et al. (2004) Am. J. Physiol. Renal Physiol. 287, F713-F721.
7. Al-Khalili, L. et al. (2004) J. Biol. Chem. 279, 25211-25218.
8. Tian, J. et al. (2006) Mol. Biol. Cell 17, 317-326.
9. Liang, M. et al. (2006) J. Biol. Chem. 281, 19709-19719.

• Béliveau, F. et al. (2011) J Biol Chem 286, 29035-43. Applications: Western Blotting
• Kashatus, D.F. et al. (2011) Nat Cell Biol 13, 1108-15. Applications: Western Blotting
• Ramos, K.M. et al. (2010) Neuroscience 169, 1888-900. Applications: Western Blotting

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• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

For Research Use Only. Not For Use In Diagnostic Procedures.