Rabbit anti-Rat Aquaporin 2 (Ser261) Polyclonal Antibody | anti-AQP2 antibody

Anti-Phospho-Ser261 Aquaporin 2

Cat. Num. AAA502006

• Gene Names: Aqp2; AQP-2; aquaporin-2
• Reactivity: Rat
• Applications: Western Blot, Immunohistochemistry
• Purity: Affinity Purified (Prepared from rabbit serum by affinity purification via sequential chromatography on phospho- and dephosphopeptide affinity columns.)
• Synonyms: Aquaporin 2 (Ser261); Polyclonal Antibody; Anti-Phospho-Ser261 Aquaporin 2; anti-AQP2 antibody

• Host: Rabbit
• Reactivity: Rat
• Clonality: Polyclonal
• Specificity: Specific for ~29k AQP2 protein phosphorylated at Ser261. Also recognizes the glycosylated form of AQP2 at ~ 37k. Immunolabeling of the AQP2 band is blocked by preadsorption with the phospho-peptide used as antigen but not by the corresponding dephospho-peptide.
• Purity/Purification: Affinity Purified (Prepared from rabbit serum by affinity purification via sequential chromatography on phospho- and dephosphopeptide affinity columns.)
• Form/Format: 100 ul in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 ug per ml BSA and 50% glycerol. Adequate amount of material to conduct 10-mini Western Blots.
• Sequence Length: 271

• Applicable Applications for anti-AQP2 antibody: Western Blot (WB), Immunohistochemistry (IHC)
• Application Notes: Quality Control: Western blots performed on each lot.; WB: 1:1000; IHC: 1:100 (Hoffert et al., 2007)
• Antigen: Phosphopeptide corresponding to amino acid residues surrounding the phospho-Ser261 of rat aquaporin 2.
• Immunogen Information: Synthetic phospho-peptide corresponding to amino acid residues surrounding Ser261 conjugated to KLH
• Immunogen Species: Rat
• Reactivity Assumed Based on 100% Sequence Homology: Canine and bovine, chicken, human, mouse, non-human primate
• Species Reactivity Note: The antibody has been directly tested for reactivity in Western blots with rat tissue. It is anticipated that the antibody will react with canine, bovine, chicken, human, mouse and non-human primate tissues based on the fact that these species have 100% homology with the amino acid sequence used as antigen.
• Biological Significance: Aquaporin 2 (AQP2) is a hormonally regulated water channel located in the renal collecting duct. Mutations in the AQP2 gene cause hereditary nephrogenic diabetes insipidus in humans (Iolascon et al.,2007). A vasopressin induced cAMP increase results in the phosphorylation of AQP2 at serine-256 and its translocation from the intracellular vesicles to the apical membrane of principal cells (van Balkom et al., 2002). Recently, serine-261 has been identified as a novel phosphorylation site on AQP2 and levels of phosphorylated S261 have been shown to decrease with vasopressin treatment suggesting its involvement in vasopressin-dependent AQP2 trafficking (Hoffert et al., 2007).
• Preparation and Storage: Store at -20 degree C; stable for at least one year.

Testing Data

Related Product Information for anti-AQP2 antibody

Affinity purified rabbit polyclonal antibody

References

• van Balkom BW, Savelkoul PJ, Markovich D, Hofman E, Nielsen S, van der Sluijs P, Deen PM (2002) The role of putative phosphorylation sites in the targeting and shuttling of the aquaporin 2 water channel. J Biol Chem 277(44):41473-9.
• Ford P, Rivarola V, Chara O, Blot-Chabaud M, Cluzeaud F, Farman M, Parisi M, Capurro C (2005) Volume regulation in cortical collecting duct cells: role of AQP2. Biol Cell 97(9):687-97.
• Hoffert JD, Nielsen J,, Yu MJ,, Pisitikun T,, Schleicher SM,, Nielsen Knepper MA (2007) Dynamics of aquaporin-2 serine-261 phosphorylation in response to short-term vasopressin treatment in collecting duct.. Am J Physiol Renal Physiol 292: F691-F700..
• Iolascon A, Aglio V, Tamma G, D'Appolito M, Addabbo F, Procino G, Simonetti MC, Montini G, Gesulado L, Debler EW, Suelto M, Valenti G (2007) Characterization of two novel missense mutations in AQP2 gene causing nephrogenic diabetes insipidus. Nephron Physiol. 105(3): p33-41.
• Hoffert JD, Fenton RA, Moeller HB, Simons B, Tchapyjnikov D, McDill BW, Yu MJ, Pisitkun T, Chen F, Knepper MA. (2008) Vasopressin-stimulated increase in phosphorylation at serine 269 potentiates plasma membrane retention of aquaporin 2. J Biol Chem. 2008 Sep 5; 283(36):24617-27

NCBI and Uniprot Product Information

• NCBI GI #: 77539434
• NCBI GeneID: 25386
• NCBI Accession #: NP_037041.2
• NCBI GenBank Nucleotide #: NM_012909.2
• UniProt Accession #: P34080; A1A5L4
• Molecular Weight: 29/37
• NCBI Official Full Name: aquaporin-2
• NCBI Official Synonym Full Names: aquaporin 2 (collecting duct)
• NCBI Official Symbol: Aqp2
• NCBI Official Synonym Symbols: AQP-2; aquaporin-2
• NCBI Protein Information: aquaporin-2; AQP-CD; WCH-CD; aquaporin-CD; ADH water channel; collecting duct water channel protein; water channel protein for renal collecting duct
• UniProt Protein Name: Aquaporin-2
• UniProt Gene Name: Aqp2
• UniProt Synonym Gene Names: AQP-2; AQP-CD
• UniProt Entry Name: AQP2_RAT

NCBI Description

water channel protein involved in water reabsorption and vasopressin regulation in kidney collecting duct cells; mutation is linked to nephrogenic diabetes insipidous (NDI) [RGD, Feb 2006]

Uniprot Description

Function: Forms a water-specific channel that provides the plasma membranes of renal collecting duct with high permeability to water, thereby permitting water to move in the direction of an osmotic gradient.

Subcellular location: Apical cell membrane; Multi-pass membrane protein. Basolateral cell membrane; Multi-pass membrane protein. Cytoplasmic vesicle membrane; Multi-pass membrane protein. Golgi apparatus › trans-Golgi network membrane; Multi-pass membrane protein. Note: Shuttles from vesicles to the apical membrane. Vasopressin-regulated phosphorylation is required for translocation to the apical cell membrane. PLEKHA8/FAPP2 is required to transport AQP2 from the TGN to sites where AQP2 is phosphorylated. Ref.6

Tissue specificity: Expressed in renal collecting tubules.

Domain: Aquaporins contain two tandem repeats each containing three membrane-spanning domains and a pore-forming loop with the signature motif Asn-Pro-Ala (NPA).

Post-translational modification: Ser-256 phosphorylation is necessary and sufficient for expression at the apical membrane. Endocytosis is not phosphorylation-dependent

By similarity.

Sequence similarities: Belongs to the MIP/aquaporin (TC 1.A.8) family. [View classification]

Sequence caution: The sequence AAA41478.1 differs from that shown. Reason: Erroneous initiation.