Anti-Aquaporin 2-ATTO-550

Anti-Aquaporin 2-ATTO-550

AQP2, AQP-CD, WCH-CD
Cat #: AQP-002-AO
Sizes: 50 µl
Source: Rabbit
Type: Polyclonal
Applications: IC, IH
Reactivity: H, M, R

Anti-Aquaporin 2-ATTO-550Application 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

Immuno-colocalization of Aquaporin 2 and TRPV4 in rat kidney.Immunohistochemical staining of rat kidney sections using Anti-TRPV4 (#ACC-034) and Anti-Aquaporin 2-ATTO-550 (AQP-002-AO) antibodies. 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.

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

Alomone Labs is pleased to offer a new version of this antibody that 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 (#AQP-002-AO) has been tested in immunohistochemistry and immunocytochemistry. It is specially suited for experiments requiring simultaneous labeling of different markers.

For a list of product citations in the literature, see product citations tab below. If you know of additional relevant citations for this product, please let us know.

• Applications
• Specifications
• Scientific Background
• Citations
• Related Products

Immunohistochemistry

Expression of Aquaporin 2 in rat kidney
Immunohistochemical staining of rat kidney paraffin embedded region using Anti-Aquaporin 2-ATTO-550 (#AQP-002-AO), (1:50), (Red). Aquaporin 2 is detected in collecting ducts but not in thin segments of the loop of Henle. Nuclei are visualized with Hoechst 33342 (blue).
Immunocytochemistry
Mouse mpkCCD_c14 cells (see Mamenko, M. et al. (2016) J. Am. Soc. Nephrol. In press.).
Immunogen
Peptide (C)RQSVELHSPQSLPRGSKA, corresponding to amino acid residues 254-271 of rat Aquaporin 2 (Accession P34080).  Intracellular, C-terminus.

HomologyMouse - identical; human, sheep - 17/18 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.
LabelATTO-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.
Standard quality control of each lotWestern blot analysis (unlabeled antibody, #AQP-002), and immunohistochemistry (labeled antibody).
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.
Reconstitution50 µl double distilled water (DDW).
Antibody concentration after reconstitution1 mg/ml.
Storage after reconstitutionThe 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 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

Aquaporin 2 (AQP-2) 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 (AQP-0 to AQP-12). Little is known about the function of the two newest members, AQP-11 and AQP-12.

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 AQP-3, AQP-7, AQP-9 and AQP-10.¹

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

AQP-2 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 AQP-2 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 vasopressing receptor induces an increase in intracellular cAMP and subsequent PKA activation, which phosphorylates AQP-2. This phosphorylation causes the translocation of AQP-2 channels from intracellular vesicles to the cell membrane where it markedly increases water permeability.^{1, 3, 4}

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.
5. Published figures using this product
6. 
   Immuno-colocalization of SK3 and Aquaporin 2 in mouse kidney
   Immunohistochemical staining of mouse kidney sections using Anti-K_Ca2.3 (SK3) (N-term) antibody (#APC-025) and Anti-Aquaporin 2-ATTO-550 antibody (#AQP-002-AO). A. Aquaporin 2 staining (red). B. SK3 staining (green). C. Merge of A and B shows SK3 expression in Aquaporin 2 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
7. Human kidney sections (1:400).
   Schwaderer, A.L. et al. (2016) J. Am. Soc. Nephrol. 27, 3175.
8. Mouse kidney sections (1:200).
   Li, Y. et al. (2016) PLoS ONE 11, e0155006.
9. Mouse kidney sections (1:200).
   Berrout, J. et al. (2014) PLoS ONE 9, e95149.
10. Rat kidney sections (1:200).
    Zaika, O. et al. (2013) J. Am. Soc. Nephrol. 24, 604.
11. 
    Immunocytochemistry citations
12. Mouse mpkCCD_c14 cells.
    Mamenko, M. et al. (2016) J. Am. Soc. Nephrol. In press.
13. 
    More product citations
14. Berrout, J. et al. (2012) J. Biol. Chem. 287, 8782.
15. Jin, M. et al. (2012) Cell Calcium 51, 131.
16. Mamenko, M. et al. (2011) PLoS ONE 6, e22824.