Anti-Nicotinic Acetylcholine Receptor β2 (extracellular)-ATTO-594 Antibody

nAChRβ2, CHRNB2
    Cat #: ANC-012-AR
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)EDFDNMKKVRLP, corresponding to amino acid residues 96-107 of rat nAChRβ2 (Accession P12390). Extracellular, N-terminus.
    Accession (Uniprot) Number P12390
    Gene ID 54239
    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.
    Label ATTO-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 1 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × 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 (unlabeled antibody, #ANC-012), and immunohistochemistry (labeled antibody).
    Applications: ic, ih, lci
    Immunohistochemistry
    Expression of nicotinic acetylcholine receptor β2 in rat cerebellum
    Immunohistochemical staining of rat cerebellum using Anti-Nicotinic Acetylcholine Receptor β2 (extracellular)-ATTO-594 Antibody (#ANC-012-AR). A. Staining shows labeling of cells in the granule layer (GL) and of some Purkinje cells (arrows). B. DAPI is used as the counterstain (blue).
    Live cell imaging / Immunocytochemistry
    Expression of Nicotinic Acetylcholine Receptor β2 in rat PC12 cells
    Immunocytochemical staining of live intact rat pheochromocytoma PC12 cells. A. Cells were stained with Anti-Nicotinic Acetylcholine Receptor β2 (extracellular)-ATTO-594 Antibody (#ANC-012-AR), (1:50), (red). B. Live view of the cells. C. Merge of the two images.
    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. Blonde, A. et al. (2000) Psychopharmacology (Berlin) 149, 293.
    5. Wilens, T.E. et al. (1999) Am. J. Psychiatry 156, 1931.
    6. Tanner, C.M. et al. (2002) Neurology 58, 581.
    7. Schneider, J.S. et al. (1998) Ann. Neurol. 43, 311.
    Scientific background

    Acetylcholine, released by cholinergic neurons, activates two groups of acetylcholine receptors (AChRs); muscarinic AChRs (mAChRs) which belong to the superfamily of G-protein coupled receptors (GPCRs) and nicotinic AChRs (nAChRs) which belong to the ligand-gated ion channel superfamily.

    nAChRs also respond to nicotine, hence their name1. 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.

    While most β subunits are neuronal, the β1 subunit forms functional receptors along with other subunits in the muscle3. The diversity of these receptors and their functional organization gives rise to unique properties and functions. The α4β2 receptor composition makes up a high affinity nicotinic receptor. In fact, its upregulation (mainly expressed by the increase of functional receptors at the membrane and not expression per se) is responsible for the increased appearance of binding sites following nicotine administration1,3.

    Animal studies have shown that nAChR-related mechanisms are involved in attention function4. Indeed α4β2 nAChR seems to also be involved in attention-deficit hyperactivity disorder (ADHD), a disease distinguished by a lack of attention, distractibility and hyperactivity3. The α4β2 and α7 nAChRs appear to be critical in rats for attention and working memory. Also, a α4β2 specific agonist was shown to reduce impulsivity, hyperactivity and attention deficits in adults with ADHD5. This same receptor subtype may also be involved in Parkinson’s disease (PD) as smoking and α4β2 nAChR agonists show beneficial effects in PD6,7.

    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:

    Immuno-colocalization of nAChRα4 and nAChRβ2 in mouse brainImmunohistochemical staining of perfusion-fixed frozen mouse parietal cortex sections using Anti-Nicotinic Acetylcholine Receptor α4 (extracellular) Antibody (#ANC-004), (1:300) and Anti-Nicotinic Acetylcholine Receptor β2 (extracellular)-ATTO-594 Antibody (#ANC-012-AR), (1:60). A. nAChRα4 staining (green). B. Same section stained for nAChRβ2 (red). C. Merge of the two images reveals several cells expressing both receptors (arrows). Nuclei are stained with DAPI (blue).

    Last update: 22/10/2018

    Anti-Nicotinic Acetylcholine Receptor β2 (extracellular) Antibody (#ANC-012) is a highly specific antibody directed against an epitope the rat protein. The antibody can be used in western blot, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize nAChRβ2 from mouse, rat, and human samples.

    Anti-Nicotinic Acetylcholine Receptor β2 (extracellular)-ATTO-594 Antibody (#ANC-012-AR) is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-Nicotinic Acetylcholine Receptor β2 (extracellular)-ATTO-594 Antibody is especially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use