Anti-Orexin Receptor 1-ATTO-488 Antibody

OX1R, Hypocretin receptor type 1, HCRTR1
    Cat #: AOR-001-AG
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)RNWKRPSEQLEAQH, corresponding to amino acid residues 256-269 of rat OX1R (Accession P56718). 3rd intracellular loop.
    Accession (Uniprot) Number P56718
    Gene ID 25593
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human - 9/14 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-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,#AOR-001), and immunohistochemistry (labeled antibody).
    Applications: ih
    May also work in: ic
    Immunohistochemistry
    Immuno-colocalization of OX1R and Parathyroid Hormone 1 Receptor in rat ventromedial hypothalamus
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Orexin Receptor 1-ATTO-488 Antibody (#AOR-001-AG), (1:60) and Anti-Parathyroid Hormone 1 Receptor (extracellular) Antibody (#APR-051), (1:60). A. Sections were incubated Anti-Parathyroid Hormone 1 Receptor (extracellular) Antibody, followed by donkey-anti-rabbit-Cy3 (red). B. The same sections were incubated with Anti-Orexin Receptor 1-ATTO-488 Antibody, (green). C. Merge of A and B shows staining of OX1R in nerve fibers (vertical arrows) while Parathyroid Hormone 1 Receptor is apparent on cell bodies (horizontal arrows). Cell nuclei are stained with DAPI (blue).
    References
    1. Sakurai, T. et al. (1998) Cell 92, 573.
    2. Kukkonen, J.P. et al. (2002) Am. J. Physiol. Cell. Physiol. 283, C1567.
    3. Kirchgessner, A.L. (2002) Endocr. Rev. 23, 1.
    4. Sakurai, T. (2007) Nat. Rev. Neurosci. 8, 171.
    Scientific background

    Orexin Receptor 1 (OX1R) (also known as hypocretin receptor 1) is one of two receptors that recognize the peptide neurotransmitters orexin A and orexin B.1 Orexin A and B are 33 and 28 amino acids in length, respectively, and are derived from a common precursor termed prepro-orexin.

    OX1R binds orexin A with greater affinity than orexin B (a one order of magnitude difference), while OX2R binds both ligands with similar affinities.2,3

    Both OX1R and OX2R belong to the 7-transmembrane domain, G protein-coupled receptor (GPCR) superfamily.
    OX1R is thought to transmit signals through the Ga11 class of G proteins, resulting in the activation of phospholipase C with subsequent triggering of the phosphatidylinositol cascade and an influx of extracellular Ca2+, probably through transient receptor potential (TRP) channels.2,3

    The physiological functions of the orexin system (OX1R, OX2R, and their ligands) have been a matter of intense research in the last few years.

    OX1R is expressed in both the central nervous system and peripheral locations such as gastrointestinal tissues, pancreas, and testis.2 It appears to be involved in the regulation of feeding behavior in rodents since an OX1R antagonist is able to inhibit baseline feeding.2,3

    The orexin system has been shown to be involved in the regulation of sleep and wakefulness states and OX1R knockout mice show defects in the regulation of rapid eye movement (REM) sleep, among other phenotypic alterations.4 In addition, the orexin system is involved in regulating autonomic functions such as blood pressure and heart rate, as well as in mechanisms that regulate the reward response in the brain.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
    Last update: 11/12/2018

    Anti-Orexin Receptor 1 Antibody (#AOR-001) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunocytochemistry, and immunohistochemistry applications. It has been designed to recognize OX1R from rat, mouse, and human samples.

    Anti-Orexin Receptor 1-ATTO-488 Antibody (#AOR-001-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-Orexin Receptor 1-ATTO-488 Antibody is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
    Citations
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