Anti-GABA(A) γ2 Receptor (extracellular) Antibody

GABRG2
    Cat #: AGA-005
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide QKSDDDYEDYASNKT(C), corresponding to amino acid residues 39-53 of rat GABA(A) γ2 Receptor (Accession number P18508). Extracellular, N-terminus.
    Accession (Uniprot) Number P18508
    Gene ID 29709
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Human, mouse, bovine - identical; chick - 13/15 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, 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 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, ih, lci, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat brain membranes:
    1. Anti-GABA(A) γ2 Receptor (extracellular) Antibody (#AGA-005), (1:200).
    2. Anti-GABA(A) γ2 Receptor (extracellular) Antibody, preincubated with the control peptide antigen.
    Transfected cells with human GABA(A) γ2 (Kang, J.Q. et al. (2010) J. Neurosci. 30, 13895.).
    Immunohistochemistry
    Rat brain formalin-fixed frozen section.
    Immunocytochemistry
    Rat adrenal medullary cells (1:100) (Matsuoka, H. et al. (2008) J. Physiol. 586, 4825.).
    Live cell imaging / Immunocytochemistry
    Mouse spinal cord neurons (1:100) (Shrivastava, A.N. et al. (2011) J. Biol. Chem. 286, 14455.).
    Mouse hippocampal neurons (1:100) (Chou, W.H. et al. (2010) J. Neurosci. 30, 13955.).
    References
    1. Mihic, S.J. et al. (1994) J.Neurotransmit. Rev. 21, 127.
    2. Whiting, P.J. et al. (1999) Neurochem. Int. 34, 379.
    3. Ashcroft, F.M. (2000) Ion Channels and Disease Ed 1.
    4. Wallace, R.Het al. (2001) Nat. Genet. 28, 49.
    5. Baulac, S. et al. (2001) Nat. Genet. 28, 46.
    Scientific background

    The neurotransmitter GABA (γ-aminobutyric acid) inhibits the activity of signal-receiving neurons by interacting with the GABAA receptor on these cells.1 There are two major types of GABA receptors: the ionotropic GABAA (GABAA R) and the metabotropic GABAB receptors.  

    GABAA R belongs to the ligand gated ion channel superfamily.1,2 

    It is a heteropentamer, with all of its five subunits contributing to the pore formation. To date, eight subunit isoforms were cloned: α, β, γ, δ, ε, π, θ, and ρ.1

    The native GABAA receptor, in most cases, consists of 2α, 2β and 1γ subunit. Three γ subunits genes have been identified in mammals. 

    The binding of GABA to its GABAA receptor results in conformational changes that open a Cl- channel, producing an increase in membrane conductance, resulting in inhibition of neural activity.2,3 

    Recently, a genetic linkage between familial epilepsy syndrome and mutations in the γ2 subunit of the GABAA receptor have been demonstrated.4,5

    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: TNF-α-induced downregulation of surface GABA(A) γ2 Receptor.Immunocytochemical staining of rat living hippocampal neurons. A. Extracellular staining of cells using Anti-GABA(A) γ2 Receptor (extracellular) Antibody (#AGA-005), (1:100). Representative images from time course experiments with cultured neurons undergoing TNF-α treatment ranging from 10 min to 6 h in duration. Results show a rapid downregulation in surface levels of the γ2 subunit in response to TNF-α. B. Quantification data of surface γ2 cluster area, over the 6 h time course of TNF-α treatment (= 120 –158 images per time point; one-way ANOVA, Tukey’s post hoc test, **< 0.01, ***< 0.001).Adapted from Pribiag, H. and Stellwagen, D. (2013) with permission of the Society for Neuroscience.
    Last update: 07/11/2018

    Anti-GABA(A) γ2 Receptor (extracellular) Antibody (#AGA-005) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunohistochemistry, and immunocytochemistry applications. The antibody recognizes an extracellular epitope and is thus highly suited for live cell imaging. It has been designed to recognize GABA(A) γ2 from human, rat, and mouse samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Mouse brain lysate (1:300).
      Du, Z. et al. (2016) Neuroscience 329, 363.
    2. HEK 293 transfected cells.
      Wang, J. et al. (2016) Sci. Rep. 6, 35294.
    3. HEK 293T transfected cells (1:100).
      Warner, T.A. et al. (2016) Hum. Mol. Genet. 25, 3192.
    4. Mouse brain lysate.
      Lo, W.Y. et al. (2014) Neurochem. Res. 39, 1088.
    5. Mouse brain lysate (1:200).
      Vlachos, A. et al. (2013) Cereb. Cortex 23, 2700.
    Live cell imaging citations
    1. Rat living primary hippocampal neurons (1:100).
      Levi, S. et al. (2015) Neuropharmacology 88, 199.
    2. Rat living hippocampal neurons (1:100).
      Pribiag, H. and Stellwagen, D. (2013) J. Neurosci. 33, 15879.
    3. Mouse spinal cord neurons (1:100).
      Shrivastava, A.N. et al. (2011) J. Biol. Chem. 286, 14455.
    4. Mouse hippocampal neurons (1:100).
      Chou, W.H. et al. (2010) J. Neurosci. 30, 13955.
    Immunohistochemistry citations
    1. Mouse spinal cord sections.
      Cantaut-Belarif, Y. et al. (2017) J. Cell Biol. 216, 2979.
    Immunocytochemistry citations
    1. Mouse primary spinal cord neurons.
      Cantaut-Belarif, Y. et al. (2017) J. Cell. Biol. 216, 2979.
    2. Rat granule cells.
      Bazzurro, V. et al. (2018) J. Mol. Neurosci. 64, 312.
    3. Rat adrenal medullary cells (1:100).
      Matsuoka, H. et al. (2008) J. Physiol. 586, 4825.
    More product citations
    1. Yue, J. et al. (2018) Brain Res. Bull. 137, 156.
    2. de Luca, E. et al. (2017) Neuron 95, 63.
    3. Kang, J.Q. et al. (2010) J. Neurosci. 30, 13895.
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