Anti-Aquaporin 2-ATTO-550 Antibody

AQP2, AQP-CD, WCH-CD
    Cat #: AQP-002-AO
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)RQSVELHSPQSLPRGSKA, corresponding to amino acid residues 254-271 of rat AQP2 (Accession P34080). Intracellular, C-terminus.
    Accession (Uniprot) Number P34080
    Gene ID 25386
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human, sheep - 17/18 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Label ATTO-550. Maximum absorption 554 nm; maximum fluorescence 576 nm. The fluorescence is excited most efficiently in the 540 - 565 nm range. This label is related to the dye Rhodamine 6G and can be used with filters used to detect Rhodamine.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 1 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × g 5 min).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis (unlabeled antibody, #AQP-002), and immunohistochemistry (labeled antibody).
    Applications: ic, ih
    Immunohistochemistry
    Expression of Aquaporin 2 in rat kidney
    Immunohistochemical staining of rat kidney paraffin embedded region using Anti-Aquaporin 2-ATTO-550 Antibody (#AQP-002-AO), (1:50), (Red). AQP2 is detected in collecting ducts but not in thin segments of the loop of Henle. Nuclei are visualized with Hoechst 33342 (blue).
    Immunocytochemistry
    Mouse mpkCCDc14 cells (Mamenko, M. et al. (2016) J. Am. Soc. Nephrol. 27, 2035.).
    References
    1. King, L.S. et al. (2004) Nat. Rev. Mol. Cell Biol. 5, 687
    2. Deen, P.M. et al. (1994) Science 264, 92.
    3. Nielsen, S. et al. (2002) Physiol. Rev. 82, 205.
    4. Robben, J.H. et al. (2006) Am. J. Physiol. 291, F257.
    Scientific background

    Aquaporin 2 (AQP2) belongs to a family of membrane proteins that allow passage of water and certain other solutes through biological membranes. The family is composed of 13 members (AQP0 to AQP12). Little is known about the function of the two newest members, AQP11 and AQP12.

    The aquaporins can be divided into two functional groups based on their permability characteristics: the aquaporins that are only permeated by water and the aquaglyceroporins that are permeated by water and other small solutes such as glycerol. This last group includes AQP3, AQP7, AQP9 and AQP10.1

    The proteins present a conserved structure of six transmembrane domains with intracellular N- and C-termini. The functional channel is a tetramer but each subunit has a separate pore and therefore the functional channel unit, contains four pores.1

    AQP2 expression is largely confined to the kidney, particularly in the renal collecting duct where it performs a key role in water absorption and urine concentration. In fact, mutations in the AQP2 gene produce hereditary nephrogenic diabetes insipidus, a disorder that results in the excretion of large volumes of urine.2

    Under normal conditions, water homeostasis in the kidney is regulated through the anti-diuretic hormone vasopressin. Vasopressin is secreted from the pituitary gland and transported to the kidney through the blood where it binds to its receptor that is mainly expressed in cells of the collecting duct. The activated vasopressin receptor induces an increase in intracellular cAMP and subsequent PKA activation, which phosphorylates AQP2. This phosphorylation causes the translocation of AQP2 channels from intracellular vesicles to the cell membrane where it markedly increases water permeability.1,3,4

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, 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
    Image & Title:
    Anti-Aquaporin 2-ATTO-550 Antibody
    Immuno-colocalization of Aquaporin 2 and TRPV4 in rat kidney.Immunohistochemical staining of rat kidney sections using Anti-TRPV4 Antibody (#ACC-034) and Anti-Aquaporin 2-ATTO-550 Antibody (AQP-002-AO). Shown are representative confocal plane micrographs (axes are shown) and corresponding cross-sections (pointed by arrows) showing three-dimensional stacks of TRPV4 (green), AQP2 localization (red), and the combined image localization (yellow). A. Wild-type Sprague-Dawley (S/D) rat. B. Cyst monolayer from a PCK453 rat. DAPI staining is shown in blue.Adapted from Zaika, O. et al. (2013) J. Am. Soc. Nephrol. 24, 604. with permission of the American Society of Nephrology.
    Last update: 11/11/2018

    Anti-Aquaporin 2 Antibody (#AQP-002) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize the AQP2 channel from rat, mouse, and human samples.

    Anti-Aquaporin 2-ATTO-550 Antibody (#AQP-002-AO) is directly labeled with an ATTO-550 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-550 fluorescent label is related to the well known dye Rhodamine 6G and can be used with filters typically used to detect Rhodamine. Anti-Aquaporin 2-ATTO-550 Antibody is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
    Citations
    Published figures using this product
    Immuno-colocalization of SK3 and Aquaporin 2 in mouse kidney
    Immunohistochemical staining of mouse kidney sections using Anti-KCNN3 (KCa2.3, SK3) (N-term) Antibody (#APC-025) and Anti-Aquaporin 2-ATTO-550 Antibody (#AQP-002-AO). A. AQP2 staining (red). B. SK3 staining (green). C. Merge of A and B shows SK3 expression in AQP2 positive tubules (orange).
    Adapted from Berrout, J. et al. (2014) PLoS ONE 9, e95149. with kind permission of Prof. O'Neil, R.G., Dpt. for Integrative Biology, the University of Texas Health Science Center Medical School, Houston, Texas, U.S.A.
    Immunohistochemistry citations
    1. Human kidney sections (1:400).
      Schwaderer, A.L. et al. (2016) J. Am. Soc. Nephrol. 27, 3175.
    2. Mouse kidney sections (1:200).
      Li, Y. et al. (2016) PLoS ONE 11, e0155006.
    3. Mouse kidney sections (1:200).
      Berrout, J. et al. (2014) PLoS ONE 9, e95149.
    4. Rat kidney sections (1:200).
      Zaika, O. et al. (2013) J. Am. Soc. Nephrol. 24, 604.
    5. Mouse kidney sections. Also tested in TRPV4-/- mice.
      Berrout, J. et al. (2012) J. Biol. Chem. 287, 8782.

    Immunocytochemistry citations
    1. Mouse mpkCCDc14 cells.
      Mamenko, M. et al. (2016) J. Am. Soc. Nephrol. 27, 2035.
    More product citations
    1. Jin, M. et al. (2012) Cell Calcium 51, 131.
    2. Mamenko, M. et al. (2011) PLoS ONE 6, e22824.
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