Anti-GABA(A) α3 Receptor (extracellular) Antibody

γ-Aminobutyric acid receptor type A subunit α3, GABRA3
    Cat #: AGA-003
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide QGESRRQEPGDFVKQ(C), corresponding to amino acid residues 29-43 of human GABRA3 (Accession P34903). Extracellular, N-terminus.
    Accession (Uniprot) Number P34903
    Gene ID 2556
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse - identical; bovine - 14/15 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, 5% sucrose, 0.025% 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 0.75 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 -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ih, ip, lci, wb
    May also work in: ifc
    Western blot
    Western blot analysis of rat brain membranes:
    1. Anti-GABA(A) α3 Receptor (extracellular) Antibody (#AGA-003), (1:200).
    2. Anti-GABA(A) α3 Receptor (extracellular) Antibody preincubated with the control peptide antigen.
    Immunoprecipitation
    Transfected COS-7 cells (1 µg antibody/140 µg lysate) (Zhou, C. et al. (2013) J. Biol. Chem. 288, 21458.).
    Immunohistochemistry
    Expression of GABRA3 in rat cerebellum
    Immunohistochemical staining of rat cerebellum using Anti-GABA(A) α3 Receptor (extracellular) Antibody (#AGA-003), (green), (1:100). GABRA3 is localized to the molecular layer (asterisk) and a portion of the Purkinje cell body (arrows), which is outlined by axonal staining using mouse anti-neurofilament 200 (red).
    Live cell imaging / Immunocytochemistry
    Expression of GABRA3 in rat insulinoma cell line
    Immunocytochemical staining of intact living rat RIN-m cells using Anti-GABA(A) α3 Receptor (extracellular) Antibody (#AGA-003), (1:50) followed by goat anti-rabbit-AlexaFluor-550 secondary antibody. Extracellular staining (red) merged with live view of the cells.
    References
    1. Mihic, S.J. and Harris, R.A. (1997) Alcohol Health Res. World 21, 127.
    2. Whiting, P.J. (1999) Neurochem. Int. 34, 387.
    3. Fuchs, K. and Celepirovic, N. (2002) J. Neurochem82, 1512.
    Scientific background

    The neurotransmitter GABA (γ-aminobutiric acid) inhibits the activity of signal-receiving neurons by interacting with the GABAA receptor on these cells.1 Binding of GABA to its GABAA receptor results in conformational changes that open a Cl- channel, producing an increase in membrane conductance that results in inhibition of neural activity.2

    There are two major types of GABA receptors: the ionotropic GABAA receptors (GABAAR) and the metabotropic GABAB receptors (GABABR). GABAARs belong to the ligand-gated ion channel superfamily.1,2

    GABAARs are heteropentamers, in which all five subunits contribute to formation of the pore. Eight subunit isoforms have been cloned: α, β, γ, δ, ε, π, θ, and ρ.Six α subunits isoforms (α1-α6) have been shown to exist in mammals.
    In most cases, the native GABAA receptors consist of 2α, 2β, and 1γ subunits. 

    The α3-subunit is highly expressed during development (along with α2 and α5) and then declines in adulthood, where the α1-subunit becomes predominant. The failure to complete this switch could be a major predispositional factor in the development of temporal lobe epilepsy.3

    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: Anti-GABA(A) α3 Receptor (extracellular) Antibody
    GABA(A) α3 Receptor expression increases in Heterozygous GABA(A) α1 Receptor knock out.
    Immunohistochemical staining of cortices from wild type (A and C) and Het α1 KO (B and D) mice in layers II/III (A and B) and VI (C and D) of the somatosensory cortex (white scale bar, 20 μm, n > 6) using Anti-GABA(A) α3 Receptor (extracellular) Antibody (#AGA-003). The yellow boxed areas are displayed on a magnified scale below each image (A2–D2).Adapted from Zhou, C. et al. (2013) with permission of the American Society for Biochemistry and Molecular Biology.
    Last update: 04/12/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of human GABA(A) α3 receptor. Anti-GABA(A) α3 Receptor (extracellular) Antibody (#AGA-003) can be used in western blot, immunoprecipitation, immunocytochemistry, and immunohistochemistry applications. The antibody recognizes an extracellular epitope and is highly suited to detect GABRA3 in living cells. It has been designed to recognize GABRA3 from rat, mouse, and human samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Mouse brain lysates (1:500).
      Seo, S. and Leitch, B. (2014) Epilepsia doi: 10.1111/epi.12500.
    2. Guinea pig and rat medulla.
      Inoue, M. et al. (2013) Neuroscience 253, 245.
    3. Mouse brain lysates (1:500).
      Zhou, C. et al. (2013) J. Biol. Chem. 288, 21458.
    Live cell imaging citations
    1. Rat oligodendrocytes (1:250).
      Arellano, R.O. et al. (2016) Mol. Pharmacol. 89, 63.
    Immunoprecipitation citations
    1. Transfected COS-7 cells (1 µg antibody/140 µg lysate).
      Zhou, C. et al. (2013) J. Biol. Chem. 288, 21458.
    Immunohistochemistry citations
    1. Rat brain sections (1:100).
      Arellano, R.O. et al. (2016) Mol. Pharmacol. 89, 63.
    2. Mouse brain sections (1:200).
      Seo, S. and Leitch, B. (2014) Epilepsia doi: 10.1111/epi.12500.
    3. Mouse brain sections (1:500).
      Zhou, C. et al. (2013) J. Biol. Chem. 288, 21458.
    4. Rat brain sections (1:250).
      Park, H.J. et al. (2013) J. Nucl. Med. 54, 1263.
    Immunocytochemistry citations
    1. Guinea pig chromaffin cells.
      Inoue, M. et al. (2013) Neuroscience 253, 245.
    2. Rat PC12 cells.
      Inoue, M. et al. (2013) Neuroscience 253, 245.
    More product citations
    1. Caspary, D.M. et al. (2013) Neurobiol. Aging 34, 1486.
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