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Anti-Nicotinic Acetylcholine Receptor α3 (CHRNA3) (extracellular) Antibody

nAChR α3, Neuronal acetylcholine receptor subunit alpha-3, ACHA3, PAOD2

Cat #: ANC-003
Alternative Name nAChR α3, Neuronal acetylcholine receptor subunit alpha-3, ACHA3, PAOD2
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: m, r
May also work in: h*
Immunogen
  • Peptide (C)KWKPSDYQGVEFMR, corresponding to amino acid residues 91-104 of rat nAChRα3 (Accession P04757). Extracellular, N-terminus.
Accession (Uniprot) Number P04757
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse - identical; human - 11/14 amino acid residues identical.
RRID AB_10687242.
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: if, ih, wb
May also work in: ic*, ifc*, ip*, lci*
Western blot
  • Western blot analysis of rat brain membranes:
    Western blot analysis of rat brain membranes:
    1. Anti-Nicotinic Acetylcholine Receptor α3 (CHRNA3) (extracellular) Antibody (#ANC-003), (1:200).
    2. Anti-Nicotinic Acetylcholine Receptor α3 (CHRNA3) (extracellular) Antibody, preincubated with Nicotinic Acetylcholine Receptor α3/CHRNA3 (extracellular) Blocking Peptide (#BLP-NC003).
Immunohistochemistry
  • Expression of Nicotinic acetylcholine receptor α3 in rat neocortex
    Expression of Nicotinic acetylcholine receptor α3 in rat neocortex
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections were stained using Anti-Nicotinic Acetylcholine Receptor α3 (CHRNA3) (extracellular) Antibody (#ANC-003), (1:200). A. Intense staining (green) of nAChR α3 appears in apical dendrites (thin arrows). A few cortical interneurons (thick arrow) are also stained with the antibody. B. The same section was stained also for parvalbumin (red staining). An arrow depicts a neuron expressing both nAChR α3 and parvalbumin.
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. Vernino, S. et al. (2009) Auton. Neurosci. 146, 3.
  5. Xu, W. et al. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 5746.
  6. Young, T. et al. (2008) Brain Res. 1229, 118.
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 a 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.

All α subunits are expressed in neuronal cells except for the α1 subunit which is specifically expressed in the muscle3. In the peripheral autonomic nervous system, nicotinic receptors are prominent and are responsible mediating fast synaptic transmission in all peripheral autonomic ganglia4. In this system, these channel receptors are mostly made up of two α3 subunits heteromerized with three other subunits. The α3 subunit plays a most important role in this system as knockout mice suffer from severe autonomic failure accompanied by gastrointestinal and bladder malfunctions5.

Autoimmune autonomic ganglionopathy (AAG) is a neurological disorder characterized by dysfunctions in sympathetic, parasympathetic and enteric functions. Patients suffering from the disease express high levels of self antibodies against ionotropic nicotinic receptors, namely α3 subunits4. The α3 subunit is also found in dorsal root ganglion where it is most probably involved in nociception6.

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

Alomone Labs is pleased to offer a highly specific antibody directed against an epitope located at the extracellular N-terminal domain of  the rat nAChRα3. Anti-Nicotinic Acetylcholine Receptor α3 (CHRNA3) (extracellular) Antibody (#ANC-003) can be used in western blot and immunohistochemistry applications. It has been designed to recognize nAChRα3 from mouse, rat and human samples.

For research purposes only, not for human use
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