Anti-Nicotinic Acetylcholine Receptor β3 (extracellular) Antibody

nAChRβ3, CHRNB3
    Cat #: ANC-013
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    May also work in: h
    Immunogen
    Peptide (C)NAKGMKGNRREG, corresponding to amino acid residues 211-222 of rat nAChRβ3 (Accession P12391). Extracellular, N-terminus.
    Accession (Uniprot) Number P12391
    Gene ID 171131
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human - 11/12 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.
    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: ic, lci, wb
    May also work in: ifc, ih, ip
    Western blot
    Western blot analysis of rat brain membranes:
    1. Anti-Nicotinic Acetylcholine Receptor β3 (extracellular) Antibody (#ANC-013), (1:400).
    2. Anti-Nicotinic Acetylcholine Receptor β3 (extracellular) Antibody, preincubated with the control peptide antigen.
    Live cell imaging / Immunocytochemistry
    Expression of Nicotinic Acetylcholine Receptor β3 in  live intact rat PC12 cells
    Immunocytochemical staining of live intact rat PC12 pheochromocytoma cells. A. Extracellular staining of cells with Anti-Nicotinic Acetylcholine Receptor β3 (extracellular) Antibody (#ANC-013), (1:50), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells. C. Merge image of A and B.
    References
    1. Albuquerque, E.X. et al. (2009) Physiol. Rev. 89, 73.
    2. Karlin, A. et al. (1986) Ann. NY. Acad. Sci. 463, 53.
    3. Kalamida, D. et al. (2007) FEBS J. 274, 3799.
    4. Deneris, E.S. et al. (1989) J. Biol. Chem. 264, 6268.
    5. Dash, B. and Lukas, R.J. (2012) J. Biol. Chem287, 14259.
    Scientific background

    Nicotinic Acetylcholine Receptors (nAChRs) mediate the physiological effects of exogenous nicotine. They also play critical physiological roles throughout the brain and body by mediating cholinergic excitatory neurotransmission, modulating the release of neurotransmitters, and having longer-term effects on gene expression and cellular connections1.

    nAChRs are pentameric complexes made up of combinations of a number of different nAChR subunits. To date, 17 different but related subunits of nAChRs have been identified and cloned. They consist of α subunits (α1-10), which is responsible for the binding of ligands. In fact, this subunit includes a Cys-loop in the first extracellular domain that is required for agonist binding2. The other subunits responsible for making up the active receptor are the β (β1-4), γ, δ and ε subunits3. Structurally, all subunits have the following: a conserved large extracellular N-terminal domain, 3 conserved transmembrane domains, a variable cytoplasmic loop and a fourth transmembrane domain with a short extracellular C-terminal domain.  An active nAChR is generally a heteropentamer of these various subunits organized around a central pore1. The β3 subunit cannot form functional receptors when expressed alone. It only forms operational channels when coexpressed with other functional subunit combinations, and is thus commonly termed ‘orphan’ or ‘auxiliary’ subunits3.

    β3 nAChR subunit mRNA is highly expressed in dopaminergic neurons of the substantia nigra and ventral tegmental area4.

    α6/β3 nAChRs have been implicated in dopaminergic neurotransmission, nicotine dependence, anxiety, and other important neurophysiological processes5.

    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: 30/10/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of rat Nicotinic Acetylcholine Receptor β3 (nAChRβ3). Anti-Nicotinic Acetylcholine Receptor β3 (extracellular) Antibody (#ANC-013) can be used in western blot and live cell imaging applications. It has been designed to recognize nAChRβ3 from mouse, rat and human samples.

    For research purposes only, not for human use