Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody

AT2 Receptor, AGTR2
    Cat #: AAR-012-AG
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    Immunogen
    Peptide DNLNATGTNESAFNC, corresponding to amino acid residues 21-35 of rat AT2 Receptor (Accession P35351). Extracellular, N-terminus.
    Accession (Uniprot) Number P35351
    Gene ID 24182
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.025% NaN3.
    Specificity Will not recognize human AT2 Receptor.
    Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.
    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, #AAR-012), and immunohistochemistry (labeled antibody).
    Applications: ic, ih, lci
    Immunohistochemistry
    Expression of Angiotensin II Receptor Type-2 in rat brain
    Immunohistochemical staining of rat frozen free floating brain sections using Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG), (1:20). AT2 receptor (green) was detected in neurons in the vicinity of the hypothalamic paraventricular nucleus (PVN). In some neurons (thick arrow), axonal processes with varicosities were observed (thin arrow). Nuclei were visualized with DAPI counterstain (blue).
    Immuno-colocalization of AT2 Receptor and VGLUT2 in rat supraoptic hypothalamic nucleus
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG), (1:60) and Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60). A. AT2 Receptor staining (green). B. VGLUT2 staining (red). C. Merge of the two images shows a general lack of co-localization of AT2 and VGLUT2 in this part of the hypothalamus (arrow). Cell nuclei are stained with DAPI (blue).
    Live cell imaging / Immunocytochemistry
    Expression of Angiotensin II Receptor Type-2 in mouse 3T3-L1 cells
    Immunocytochemical staining of intact live mouse 3T3-L1 cells with Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG), (1:50), (green). Live view of the same field was superimposed to the fluorescent one.
    References
    1. Mukoyama, M. et al. (1993) J. Biol. Chem. 268, 24539.
    2. de Gasparo, M. et al. (2000) Pharmacol. Rev. 52, 415.
    3. Carey, R.M. and Siragy, H.M. (2003) Endocr. Rev. 24, 261.
    Scientific background

    Angiotensin II receptor type 2 or AT2 is one of the receptors that bind the octapeptide hormone Angiotensin II (Ang II).
     
    Ang II is the peptide hormone that generates most of the known effects of the renin-angiotensin system (RAS). Ang II is generated from the angiotensinogen protein by the actions of renin, angiotensin converting enzyme (ACE) and other peptidases. Ang II has a central role in cardiovascular homeostasis by regulating vasoconstriction, renal Na+ and water readsorption. In addition, Ang II induces cell growth and proliferation and has pro-inflammatory effects.
     
    Most of the physiological actions of Ang II are mediated by AT1 a member of the 7-transmembrane domain, G protein-coupled receptor (GPCR) superfamily. AT2 is also a GPCR, and is largely believed to have counter-regulatory roles to the ones exerted through the AT1 receptor. Hence, binding of Ang II to the AT2 receptor will result in growth arrest and apoptosis, vasodilatation and hypotension.
     
    Although the AT2 receptor is structurally a member of the GPCR superfamily, the signaling mechanisms elicited following AT2 receptor activation are not fully clarified. Recent evidence indicates that the AT2 receptor signals through the Giα2 and the Giα3 proteins and through the activation of phosphotyrosine phosphatases such as  SHP-1.
     
    AT2 receptor distribution is more restricted than that of the AT1 receptor. The highest expression levels of AT2 have been found in the fetus, which is followed by a marked decrease shortly after birth. Nevertheless, significant AT2 receptor expression can be detected in the heart, kidney, vascular endothelial cells and brain, among others.

    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-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody
    Immuno-colocalization of Melatonin Receptor Type 2 and AT2 Receptor in rat brain
    Immunohistochemical staining of perfusion-fixed frozen brain sections using Anti-Melatonin Receptor Type 2 Antibody (#AMR-032), (1:600) and Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG), (1:100). A. Melatonin Receptor Type 2 staining (red) (arrows). B. The same section labeled for Angiotensin II Receptor Type-2 (green). C. Merge of the two images suggests considerable co-localization in the paraventricular nucleus (arrows). For orientation, note localization with respect to 3rd ventricle (3rd V).

    Last update: 23/10/2018

    Anti-Angiotensin II Receptor Type-2 (extracellular) Antibody (#AAR-012) is a highly specific antibody directed against an extracellular epitope of the rat protein. The antibody can be used in western blot, immunocytochemistry and immunohistochemistry applications. It has been designed to recognize AT2 receptor from rat and mouse samples. The antibody won’t recognize human AT2 receptor.

    Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG) is directly labeled with an ATTO-488 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-488 label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody has been tested in live cell imaging and immunohistochemical applications and is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use