Anti-CHRM2 Antibody

Muscarinic acetylcholine receptor M2, Cholinergic receptor muscarinic 2, mAChR M2
    Cat #: AMR-002
    Alternative Name Muscarinic acetylcholine receptor M2, Cholinergic receptor muscarinic 2, mAChR M2
  • KO Validated
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • GST fusion protein with the sequence VANQDPVSPSLVQGRIVKPN NNNMPSSDDGLEHNKIQNGKAPRDPVTENCVQGEEKESSNDSTSV SAVASNMRDDEITQDENTVSTSLGHSKDENSKQTCIRIGTKTPKS DSCTPTNTTVEVVGSSGQNGDE, corresponding to amino acid residues 225-356 of human M2 (Accession P08172). 3rd intracellular loop.
    • Anti-CHRM2 Antibody
    Accession (Uniprot) Number P08172
    Gene ID 1129
    Peptide confirmation Confirmed by DNA sequence and SDS-PAGE.
    Homology Chimpanzee, gorilla - identical; orangutan - 130/132 amino acid residues identical; pig - 122/132 amino acid residues identical; rat - 118/132 amino acid residues identical; mouse - 117/132 amino acid residues identical; dog - 122/132 amino acid residues identical.
    RRID AB_2039995.
    Purity The serum was depleted of anti-GST antibodies by affinity chromatography on immobilized GST and then the IgG fraction was 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).
    Negative 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.
    Negative control antigen reconstitution 100 μl PBS.
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 3 µg fusion protein per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, if, ih, ip, wb
    May also work in: ifc*
    Western blot
    • Anti-CHRM2 Antibody
      Western blot analysis of rat brain membranes:
      1. Anti-CHRM2 Antibody (#AMR-002), (1:200).
      2. Anti-CHRM2 Antibody, preincubated with the negative control antigen.
    Immunoprecipitation
    • Xenopus oocyte membranes (Vorobiov, D. et al. (2000) J. Biol. Chem. 275, 4166.).
    Immunohistochemistry
    • Anti-CHRM2 Antibody
      Expression of Muscarinic acetylcholine receptor M2 in mouse parieto-temporal cortex sections
      Immunohistochemical staining of mouse parieto-temporal cortex frozen sections (non-consecutive) using Anti-GIRK2 (Kir3.2) Antibody (#APC-006), (1:100) and Anti-CHRM2 Antibody (#AMR-002), (1:100). mAChR M2 staining (red) was dense in layer IV, with fibers climbing to layers II-III. Kir3.2 K+ channel staining (green) was dense in layers IV and I. Overlapping expression of Kir3.2 channel and mAChR M2 is seen in cortical layers.
    • Human colon (1:50) (Harrington, A.M. et al. (2010) Neurogastroenterol. Motil. 22, 999.).
    Immunocytochemistry
    • Mouse HL-1 cells (1:500) (Nobles, M. et al. (2010) Pflugers Arch. 460, 99.).
    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. Ferreira, A.R. et al. (2003) Pharmacol. Biochem. Behav. 74, 411.
    4. Van der Zee, E.A. and Luiten, P.G. (1999) Prog. Neurol. 58, 409.
    5. Tata, A.M. et al. (1999) Brain Res. 824, 63.
    6. Shapiro, M.S. et al. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 10899.
    7. Jin, W. and Lu, Z. (1998) Biochemistry 37, 13291.
    Scientific background

    The action of the neurotransmitter acetylcholine is mediated through two types of receptors, the ionotropic nicotinic receptors and the metabotropic muscarinic receptors. The muscarinic receptors belong to the superfamily of 7-transmembrane G-protein coupled receptors. Five subtypes of muscarinic receptors have been cloned and are named M1-M5.1-2

    The muscarinic receptors are widely distributed throughout the body but are predominantly expressed in the parasympathetic nervous system and exert both excitatory and inhibitory control over central and peripheral tissues.1-2

    Muscarinic receptors participate in a number of physiological functions such as regulation of heart rate, muscle contraction, cognition, sensory processing, and motor control.1 They also participate in learning and memory processing.3-4

    The M2 receptor is considered to be the predominant muscarinic receptor subtype that is expressed in cardiac muscle.5

    The M2 and M4 receptors mediate Ca2+ channel inhibition and Kir3 K+ channel activation by directly binding the Gβγ subunit to the channel.6,7 Stimulation of the M2 receptor by acetylcholine in the heart results in activation of the Kir3.1/Kir3.4 channels causing a slowing in heart beat.7

    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: 24/01/2020

    Anti-CHRM2 Antibody (#AMR-002) is a highly specific antibody directed against an epitope of the human M2 muscarinic receptor. The antibody can be used in western blot, immunoprecipitation, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize M2 from mouse, rat, and human samples.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    KO validation citations
    1. Immunohistochemical staining of mouse brain sections. Tested in M2R-/- mice.
      Garzon, M. and Pickel, V.M. (2006) J. Comp. Neurol. 498, 821.
    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 colon lysate.
      Kim, J.E. et al. (2017) BMC Gastroenterol. 17, 21.
    4. Rat colon lysate.
      Kim, J.E. et al. (2016) PLoS ONE 11, e0161144.
    5. Rat bladder lysate (1:1000).
      Lee, W.C. et al. (2016) Sci. Rep. 6, 34669.
    6. Mouse ventricle lysate.
      Liu, Y. et al. (2013) J. Transl. Med. 11, 209.
    Immunoprecipitation citations
    1. Xenopus oocyte membranes.
      Vorobiov, D. et al. (2000) J. Biol. Chem. 275, 4166.
    Immunohistochemistry citations
    1. Zebrafish brain and spinal cord sections.
      Bertuzzi, M and Ampatzis, K. (2018) Sci. Rep. 8, 1988.
    2. Rat brain sections.
      Garzon, M. and Pickel, V.M. (2016) J. Comp. Neurol. 524, 3084.
    3. Human colon (1:50).
      Harrington, A.M. et al. (2010) Neurogastroenterol. Motil. 22, 999.
    4. Mouse brain sections. Also tested in M2R-/- mice.
      Garzon, M. and Pickel, V.M. (2006) J. Comp. Neurol. 498, 821.
    Immunocytochemistry citations
    1. Mouse BMVECs (brain endothelial microvascular cells), (1:100).
      Radu, B.M. et al. (2017) Sci. Rep. 7, 5083.
    2. Mouse HL-1 cells (1:500).
      Nobles, M. et al. (2010) Pflugers Arch. 460, 99.
    More product citations
    1. Gregory, K.J. et al. (2012) J. Biol. Chem. 287, 37006.
    2. Lee, K.B. et al. (2000) J. Biol. Chem. 275, 35767.
    3. Rybin, V.O. et al. (2000) J. Biol. Chem. 275, 41447.
    Shipping and Ordering information