Anti-Melatonin Receptor Type 2 Antibody

MT2, Mel1b Receptor, MTNR1B
    Cat #: AMR-032
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    Immunogen
    Peptide (C)RKAKATRKLRLRPSD, corresponding to amino acid residues 232-246 of mouse Melatonin Receptor Type 2 (Accession Q8CIQ6). 3rd intracellular loop.
    Accession (Uniprot) Number Q8CIQ6
    Gene ID Q8CIQ6
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat - 14 /15 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.
    Specificity Unlikely to recognize human samples.
    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).
    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 -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ih, wb
    May also work in: ic, ifc, ip
    Western blot
    Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain lysates:
    1,2. Anti-Melatonin Receptor Type 2 Antibody (#AMR-032), (1:200).
    3,4. Anti-Melatonin Receptor Type 2 Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Immuno-colocalization of Melatonin receptor 2 and SCN2A in rat paraventricular nucleus
    Immunohistochemical staining of perfusion-fixed frozen rat paraventricular nucleus sections using Anti-Melatonin Receptor Type 2 Antibody (#AMR-032), (1:600) and Guinea pig Anti-SCN2A (NaV1.2) Antibody (#AGP-026), (1:2000). A. Melatonin Receptor Type 2 staining (red) (arrows). B. The same section labeled for SCN2A (green). C. Merge of A and B demonstrates partial co-localization of Melatonin Receptor Type 2 and SCN2A in the paraventricular nucleus (PVN). For orientation, note location with respect to the third ventricle (3rd V). Cell nuclei were stained with DAPI (blue).
    Immuno-colocalization of Melatonin Receptor Type 2 and Angiotensin II Receptor Type 2 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).
    Expression of Melatonin receptor type 2 in rat supraoptic nucleus
    Immunohistochemical staining of perfusion-fixed frozen brain sections using Anti-Melatonin Receptor Type 2 Antibody (#AMR-032), (1:600), (red). Melatonin Receptor Type 2 is expressed discretely in the supraoptic  nucleus (SON), adjacent to the optric tract (OT). DAPI is used as the counterstain (blue).
    References
    1. Luchetti, F. et al. (2010) FASEB J. 24, 3603.
    2. Zawilska, J.B. et al. (2009) Pharmacol. Rep. 61, 383.
    3. Hardeland, R. (2009) Biofactors 35, 183.
    4. Altun, A. and Ugur-Altun, B. (2007) Int. J. Clin. Pract. 61, 835.
    Scientific background

    Melatonin (N-acetyl-5-methoxytryptamine) is a product of tryptophan metabolism. It is synthesized in the pineal gland and is secreted to control the circadian rhythm1-3. Melatonin is also synthesized in the gastrointestinal tract, retina, skin and other tissues where it acts in a autocrine or paracrine manner. The role of melatonin in these tissues is independent of its role in the circadian rhythm, where it plays a role in energy metabolism, physiological growth, differentiation and responsiveness in stress stimuli1. The pleiotropic effects of melatonin have given rise to various therapeutic possibilities for this molecule. For example; anti-stress, sexual dysfunction, obesity, gallbladder stones1,4. To date, the only therapeutic uses for melatonin remain to treat sleep disorders, depression, migraine and headaches1.

    Melatonin exerts its effects through two G-protein coupled receptors (GPCRs); melatonin receptor type 1 and melatonin receptor type 2 (MT1 and MT2). Like all GPCRs, they have seven transmembrane domains and extracellular N-terminal and cytoplasmic C-terminal tails. The binding of melatonin to either receptor activates Gi, thereby activating PLC, thus increasing intracellular Ca2+ levels. Both receptors structurally bind melatonin in the same manner, although MT2 displays a much higher affinity for the hormone. Just like melatonin levels are detected in many tissues, the expression patterns of the two receptors are also quite broad. For example, MT1 is detected in the brain, retina and kidneys and MT2 is expressed in brain and in the retina1.

    MT1 is involved in sleep regulation and might also have effects on peripheral vasoconstriction. MT2 may play an important physiological role in the retina and might regulate body temperature1.

    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:
    Expression of MT2 in rat retina.Immunohistochemical staining of rat retinal sections using Anti-Melatonin Receptor Type 2 Antibody (#AMR-032). MT2 staining (green) is restricted to the cytoplasm of retinal ganglion cells.Adapted from Sheng, W.L. et al. (2015) PLoS ONE 10, e0117967. with permission of PLoS.
    Last update: 01/11/2018

    Anti-Melatonin Receptor Type 2 Antibody (#AMR-032) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize MT2 from rat and mouse samples. The antibody is unlikely to recognize the receptor in human samples.

    For research purposes only, not for human use
    Citations
    Published figures using this product
    Expression of MT1 and MT2 receptors in rat retina.
    A1-E2: Western blot analysis: of rat retina lysate: A1. Anti-Melatonin Receptor Type 1 Antibody (#AMR-031). A2. Anti-Melatonin Receptor Type 1 Antibody preincubated with the control peptide antigen. E1. Anti-Melatonin Receptor Type 2 Antibody (#AMR-032). E2. Anti-Melatonin Receptor Type 2 Antibody preincubated with the control peptide antigen. B1-H2: Immunohistochemical staining of rat retina sections using Anti-Melatonin Receptor Type 1 Antibody (#AMR-031) (B2, B3, C2, C3) and Anti-Melatonin Receptor Type 2 Antibody (#AMR-032) (F2, F3, G2, G3). Melatonin receptor expression (green) coincides with that of melanopsin in ipRGCs (red). Note that detection of both receptors is abolished when the antibody is preincubated with the control peptide antigen (panels D2 and H2).
    Adapted from Sheng, W.L. et al. (2015) with kind permission of Weng, S.J. Institute of Neurobiology, Institutes of Brain Science, State Key Laboratory of Medical Neurobiology and Collaborative Innovation Center for Brain Science, Fudan University, Shanghai, China.
    Immunohistochemistry citations
    1. Mouse cornea sections (1:500).
      Baba, K. et al. (2015) Invest. Ophtalmol. Vis. Sci. 56, 4753.
    2. Rat retina sections.
      Sheng, W.L. et al. (2015) PLoS ONE 10, e0117967.
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