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Anti-CHRM3 Antibody

Cholinergic muscarinic receptor 3, Muscarinic acetylcholine receptor M3, M3 mAChR

Cat #: AMR-006
Alternative Name Cholinergic muscarinic receptor 3, Muscarinic acetylcholine receptor M3, M3 mAChR
  • KO Validated
  • Lyophilized Powder yes
    Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • Peptide (C)TLAKRFALKTRSQITKRKR, corresponding to amino residues 461-479 of rat M3 (Accession P08483). 3rd intracellular loop.
    Accession (Uniprot) Number P08483
    Gene ID 24260
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Human, mouse, bovine, pig - identical.
    RRID AB_2039997.
    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, ip, wb
    May also work in: ifc*
    Western blot
    • Western blot analysis of rat brain lysate:
      Western blot analysis of rat brain lysate:
      1. Anti-CHRM3 Antibody (#AMR-006), (1:200). 
      2. Anti-CHRM3 Antibody, preincubated with CHRM3 Blocking Peptide (#BLP-MR006).
    Immunoprecipitation
    • Rat heart lysate (3 μg) (Yue, P. et al. (2006) Cell. Physiol. Biochem. 17, 129.).
    Immunohistochemistry
    • Human colon (1:50) (Harrington, A.M. et al. (2010) Neurogastroenterol. Motil. 22, 999.).
    Immunocytochemistry
    • Rat primary astrocytes (1:400) (Pap, P. et al. (2009) Hearing Res. 255, 73.).
    References
    1. Felder, C.C. et al. (2000) J. Med. Chem. 43, 4333.
    2. Forsythe, S.M. et al. (2002) Am. J. Respir. Cell. Mol. Biol. 26, 298.
    3. Hulme, E. et al. (1990) Ann. Rev. Pharmac. Toxicol. 30, 633.
    4. Ferreira, A.R. et al. (2003) Pharmacol. Biochem. Behav. 74, 411.
    5. Van der Zee, E.A. et al. (1999) Prog. Neurol. 58, 409.
    6. Levey, A.I. et al. (1991) J. Neurosci. 11, 3218.
    7. Wang, J. et al. (2000) Am. J. Physiol. Gastrointest. Liver Physiol. 279, G1059.
    8. Billington, C.K. et al. (2002) Am. J. Respir. Cell. Mol. Biol. 26, 269.
    9. Ehlert, F.J. (2003) Life. Sci. 74, 355.
    10. Budd, S.C. et al. (2003) J. Biol. Chem. 278, 19565.
    11. Frucht, H. et al. (1999) Clin. Cancer. Res. 5, 2532.
    12. Rayford, W. et al. (1997) Prostate. 30, 160.
    13. Ukegawa, J.I. et al. (2003) J. Cancer. Res. Clin. Oncol. 129, 272.
    14. Shafer, S.H. et al. (2004) Mol. Pharmacol. 65, 1080.
    Scientific background

    The action of the neurotransmitter acetylcholine is mediated through two types of receptors, the ionotrophic nicotinic receptors and the metabotrophic muscarinic receptors. The muscarinic receptors belong to the superfamily of 7-TM G-protein-coupled receptors. Five subtypes of muscarinic receptors have been cloned and named m1-m5.1-2

    The muscarinic receptors are widely distributed throughout the body, but are predominantly  expressed within the parasympathetic nervous system and exerts both excitatory and inhibitory control over central and peripheral tissues.1-2 The m3 receptor is expressed in brain, heart, urinary bladder, salivary gland and airway smooth muscle.3

    Muscarinic receptors participate in a number of physiologic functions such as regulation of heart rate,  muscle contraction, cognition, sensory processing and motor control.1 They also participate in learning and memory processing.4-5 The m1 receptors are the most abundant muscarinic subtype in the cortex and striatum. m1 receptors were also localized in the myenteric plexus where they function as autoreceptors to enhance the release of Ach from the nerves.6-7 m2, found in heart, is coupled to and mediates the slowing of heart rate in response to acetylcholine.3,8 m3 controls smooth muscle contraction and its stimulation causes secretion from glandular tissues.9

    Involvement of muscarinic receptors in apoptosis and cell growth has been reported.10-14  m1, m3 and m5, which are coupled to Gq/11 proteins, can protect cells from undergoing apoptosis induced by DNA damage. The signaling mechanism that mediates this anti-apoptotic response is still poorly understood. However, it was recently reported that a poly-basic motif in the C-terminus tail of the m3, m1 and m5 receptors is an essential element for the anti-apoptotic response of those receptors.10

    Muscarinic receptors are also capable of promoting cell proliferation in many cell types, however, the signaling pathways are yet, unclear.11-14 In the literature, m3 was reported to be involved in mitogenesis and proliferation of different cell types. Recently, it was reported that activation of the m3 muscarinic receptor induces significant proliferation of colon and prostate cancer cells.11-14

    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: 11/04/2021

    Anti-CHRM3 Antibody (#AMR-006) is a highly specific antibody directed against an epitope of the rat M3 muscarinic receptor. The antibody can be used in western blot, immunohistochemistry, immunocytochemistry, and immunoprecipitation applications. It has been designed to recognize M3 from human, mouse, and rat samples.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    KO validation citations
    1. Western blot analysis of human SH-SY5Y cells transfected with M3 receptor siRNA.
      Olianas, M.C. et al. (2014) Biochim. Biophys. Acta 1843, 3004.
    Western blot citations
    1. Mouse colon lysate
      Kim, J.E. et al. (2019) PLoS ONE 14, e0215205.
    2. Rat bladder lysate (1:1000).
      Lee, W.C. et al. (2018) Sci. Rep. 8, 5795.
    3. Rat bladder lysate.
      Lee, Y.L. et al. (2018) Sci. Rep. 8, 5358.
    4. Mouse colon epithelia lysate.
      Rottgen, T.S. et al. (2018) Am. J. Physiol. 315, C10.
    5. Rat colon lysate.
      Kim, J.E. et al. (2017) BMC Gastroenterol. 17, 21.
    6. Rat colon lysate.
      Kim, J.E. et al. (2016) PLoS ONE 11, e0161144.
    7. Rat bladder lysate (1:1000).
      Lee, W.C. et al. (2016) Sci. Rep. 6, 34669.
    8. Human SH-Sy5Y cell lysate.
      Olianas, M.C. et al. (2016) Br. J. Pharmacol. 173, 2910.
    9. Rat colon lysate.
      Kim, J.E. et al. (2016) PLoS ONE 11, e0161144.
    10. Human SH-SY5Y cell lysate.
      Olianas, M.C. et al. (2014) Biochim. Biophys. Acta 1843, 3004.
    Immunoprecipitation citations
    1. Rat heart lysate (3 μg).
      Yue, P. et al. (2006) Cell. Physiol. Biochem. 17, 129.
    Immunohistochemistry citations
    1. Human primary adenocarcinoma tumor sections.
      Cheng, K. et al. (2017) Oncotarget 8, 21106.
    2. Mouse colon sections.
      Raufman, J.P. et al. (2015) Carcinogenesis 36, 1193.
    3. Human colon (1:50).
      Harrington, A.M. et al. (2010) Neurogastroenterol. Motil. 22, 999.
    Immunocytochemistry citations
    1. Mouse BMVECs (brain endothelial microvascular cells), (1:100).
      Radu, B.M. et al. (2017) Sci. Rep. 7, 5083.
    More product citations
    1. Pap, P. et al. (2009) Hearing Res. 255, 73.
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