antibody

Na,K-ATPase alpha1, phosphorylated (Ser16)

N2298

100 ul

polyclonal

domestic rabbit

nonconjugated

phosphorylated

Na,K-ATPase alpha1, phosphorylated (Ser16)

rabbit

Pab

Phosphorylated

Antibodies

100 ul

IgG

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 alpha subunit and a beta subunit (1). Several phosphorylation sites have been identified for the alpha 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 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 alpha 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). Applications: Suitable for use in Western Blot. Other applications not tested. Recommended Dilution: Western Blot: 1:1000 Optimal dilutions to be determined by the researcher. Storage and Stability: For long-term storage, aliquot and store at -20°C. Aliquots are stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Rt

Synthetic phosphopeptide corresponding to residues surrounding Ser16 of rat Na,K-ATPase a1. Species sequence homology: mouse, bovine and porcine (100%)

Recognizes endogenous levels of rat Na,KATPase alpha1 only when phosphorylated at Ser16. The residue number, Ser16, is based on the sequence of the immature form of the protein, corresponding to Ser11 of the mature cleaved form.

Purified by Protein A and peptide affinity chromatography.

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

Not determined

Product Type: Pab Isotype: IgG Host: rabbit Source: rat Concentration: Not determined Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. Purity: Purified by Protein A and peptide affinity chromatography. Immunogen: Synthetic phosphopeptide corresponding to residues surrounding Ser16 of rat Na,K-ATPase a1. Species sequence homology: mouse, bovine and porcine (100%) Specificity: Recognizes endogenous levels of rat Na,KATPase alpha1 only when phosphorylated at Ser16. The residue number, Ser16, is based on the sequence of the immature form of the protein, corresponding to Ser11 of the mature cleaved form. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

WB

-20°C

(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.

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