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Anti-Orexin Receptor 1 Antibody

OX1R, Hypocretin receptor type 1, HCRTR1
Cat #: AOR-001
Alternative Name OX1R, Hypocretin receptor type 1, HCRTR1
Lyophilized Powder yes
Type: Polyclonal
Host: 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.
RRID AB_2040048.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 μl, 50 μl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C 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 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: ic, if, ih, wb
May also work in: ifc*, ip*
Western blot
  • Western blot analysis of mouse brain lysate:
    Western blot analysis of mouse brain lysate:
    1. Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:200).
    2. Anti-Orexin Receptor 1 Antibody, preincubated with Orexin Receptor 1 Blocking Peptide (#BLP-OR001).
  • Western blot analysis of rat brain lysate:
    Western blot analysis of rat brain lysate:
    1. Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:500).
    2. Anti-Orexin Receptor 1 Antibody, preincubated with Orexin Receptor 1 Blocking Peptide (#BLP-OR001).
  • Western blot analysis of human Colo-205 (lanes 1 and 3) and HT-29 (lanes 2 and 4) colon cancer cell lines:
    Western blot analysis of human Colo-205 (lanes 1 and 3) and HT-29 (lanes 2 and 4) colon cancer cell lines:
    1,2. Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:500).
    3,4. Anti-Orexin Receptor 1 Antibody, preincubated with Orexin Receptor 1 Blocking Peptide (#BLP-OR001).
Immunohistochemistry
  • Expression of OX1R in rat brain
    Expression of OX1R in rat brain
    Longitudinal frozen section of rat brainstem showing staining (red) in neuronal cell bodies. Slides were incubated overnight at 4°C with Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:50) followed by goat-anti-rabbit-AlexaFluor-555 secondary antibody (1:500). Hoechst 33342  is used as a counterstain (blue).
  • Expression of OX1R in rat colon
    Expression of OX1R in rat colon
    Immunohistochemical staining of paraffin-embedded longitudinal section of rat colon showing mucosa (M), submucosa (SM), and muscularis externa (ME) using Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:100). Note that the stain (red-brown color) is highly specific for absorptive cells in the superior third of the intestinal glands. Immunolabeling was detected using DAB as the chromogen and hematoxilin as the counterstain.
  • Expression of OX1R in mouse septum
    Expression of OX1R in mouse septum
    Immunohistochemical staining paraffin-fixed frozen sections using Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:50). A. OX1R (green) appears in axonal processes (right-pointing triangles). B. Parvalbumin (red) appears in septal neurons. Cell nuclei (blue) are visualized with Hoechst 33342. C. Merge of OX1R and parvalbumin suggests that orexinergic innervation covers the entire septal nucleus rather than restricted to individual neurons.
Immunocytochemistry
  • Expression of OX1R in human colon cancer cell lines
    Expression of OX1R in human colon cancer cell lines
    Immunocytochemical staining of paraformaldehyde fixed and permeabilized human Colo-205 colon cancer cells A. Cells were stained with Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:200), followed by goat-anti-rabbit-AlexaFluor-555 secondary antibody. B. Live view of the same field as in (A). C. Nuclei were visualized with the cell permeable dye Hoechst 33342 (blue staining).
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, IF- Immunofluorescence, 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: 16/08/2020

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.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
Western blot citations
  1. Rat brain lysate.
    Zhou, J.J. et al. (2015) Neuropharmacology 99, 481.
Immunohistochemistry citations
  1. Rat hypothalamus sections (1:100).
    Zhou, J.J. et al. (2015) Neuropharmacology 99, 481.
Immunocytochemistry citations
  1. Rat isolated rod bipolar cells (RBCs).
    Zhang, G. et al. (2018) Neuropharmacology 133, 38.
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