Anti-GABA(A) α1 Receptor (extracellular) Antibody

γ-Aminobutyric acid receptor type A subunit α1, GABRA1
    Cat #: AGA-001
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    May also work in: h
    Immunogen
    Peptide QPSQDELKDNTTVFTR(C), corresponding to amino acid residues 28-43 of rat GABRA1 (Accession P62813). Extracellular, N-terminus.
    Accession (Uniprot) Number P62813
    Gene ID 29705
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human, bovine - 16/17 amino acid residues identical; chicken - 14/17 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 -20°C.
    Preadsorption Control 2 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 lysates:
    1. Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001), (1:1000).
    2. Anti-GABA (A) α1 Receptor (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of GABRA1 in mouse hippocampus
    Immunohistochemical staining of mouse hippocampus using Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001). A. Distribution of GABRA1 (red). B. Distribution of glial fibrillary acidic protein (green). C. Merge of the two images indicates that distribution of GABRA1 is restricted to neurons and their processes. DAPI is used as the counterstain (blue).
    Colocalization of CaV1.2 and GABA(A) α1 Receptor in rat cerebellum
    Immunohistochemical staining of rat cerebellum using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001). A. CaV1.2 (green) is detected in the granule layer of the cerebellum (G) and in the upper molecular layer (star). B. In the same section, GABRA1 (red) is seen in the granule layer. C. Merge of the two images reveals high degree of colocalization between CaV1.2 and GABRA1 in the granule layer.
    Immunocytochemistry
    Rat adrenal medullary cells (1:100) (Matsuoka, H. et al. (2008) J. Physiol. 586, 4825.).
    Live cell imaging / Immunocytochemistry
    Expression of GABRA1 in rat PC12 cells
    Immunocytochemical staining of intact living rat pheochromocytoma PC12 cells. A. Extracellular staining of cells using Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001), (1:100) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody. B. Merge image of A and live view of the cells.
    Rat living hippocampal neurons (1:400) (Pribiag, H. and Stellwagen, D. (2013) J. Neurosci. 33, 15879.).
    References
    1. Owens, D.F. and Kriegstein, A.R. (2002) Nat. Rev. Neurosci3, 715.
    2. Whiting, P.J. (1999) Neurochem. Int. 34, 387.
    3. Mihic, S.J. and Harris, R.A. (1997) Alcohol Health Res. World 21, 127.
    4. Neelands, T.R. et al. (1999) J. Neurosci. 197057.
    5. Fuchs, K. and Celepirovic, N. (2002) J. Neurochem82, 1512.
    Scientific background

    GABA (γ-aminobutyric acid) is the major inhibitory neurotransmitter in the brain. Its production, release, reuptake, and metabolism all occur in the nervous system.1

    The GABA transmitter interacts with two major types of receptors: ionotropic GABAA receptors (GABAAR) and metabotropic receptors (GABABR). GABAARs belong to the ligand-gated ion channel superfamily.2 GABA inhibits the activity of signal-receiving neurons by interacting with the GABAA receptor on these cells.3 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

    GABAARs are heteropentamers, in which all five subunits contribute to pore formation.  To date, eight subunit isoforms have been cloned:α, β, γ, δ, ε, π, θ, and ρ.1 Six α subunit isoforms have been found to exist in mammals (α1-α6). In most cases, native GABAA receptors consists of 2α, 2β, and 1δ subunits. The α subunit is the most common and is expressed ubiquitously. It determines the affinities of GABAARs for allosteric ligands.

    Each subtype has a unique regional expression in the brain, and individual neurons often express multiple subtypes.4 The α1 subunit is highly expressed in adulthood while the α2 subunit is highly expressed very early in rat brain development. Failure to complete the normal transition between the α-subunits that are highly expressed in early development (α2, α3, and α5) and those expressed in adulthood (α1) is suggested to play a major role in the development of temporal lobe epilepsy.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:

    Anti-GABA(A) α1 Receptor (extracellular) Antibody
    High resolution immuno-scanning electron microscopy (HRSEM) of GABA(A) α1 Receptor in mouse hippocampal neurons.A. BSE (backscattered electron) analysis on mouse primary hippocampal neuron cultures labeled with Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001) followed by FluoroNanogoldTM Fab secondary antibody reveals the presence of GABRA1 gold clusters of variable size (due to gold-enhancement reaction) on both soma and neurites. B. At higher magnification, gold clusters are observed on the plasma membrane in close proximity to neurite processes covering the neuron cell body. Gold clusters are also detected at contact regions between processes (synaptic contacts).Adapted from Orlando, M. et al. (2017) Sci. Rep. 7, 13768. with permission of NATURE SPRINGER.

    Last update: 07/11/2018

    Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001) 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 highly suited to detect GABRA1 in live cells. It has been designed to recognize GABRA1 from human, rat, and mouse samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Mouse brain lysate (1:1000).
      Nys, J. et al. (2015) J. Neurosci. 35, 11174.
    2. Mouse brain lysates (1:500).
      Seo, S. and Leitch, B. (2014) Epilepsia 55, 224.
    3. Guinea pig and rat medulla.
      Inoue, M. et al. (2013) Neuroscience 253, 245.
    4. Rat brain lysates.
      Woo, J. et al. (2013) J. Cell Biol. 201, 929.
    Live cell imaging citations
    1. Mouse primary hippocampus neurons (1:30).
      Orlando, M. et al. (2017) Sci. Rep. 7, 13768.
    2. Rat oligodendrocytes (1:250).
      Arellano, R.O. et al. (2016) Mol. Pharmacol. 89, 63.
    3. Mouse living hippocampal neurons.
      Petrini, E.M. et al. (2014) Nat. Commun. 5, 3921.
    4. Rat living hippocampal neurons (1:400).
      Pribiag, H. and Stellwagen, D. (2013) J. Neurosci. 33, 15879.
    5. Rat living hippocampal neurons (1:500).
      Chaumont, S. et al. (2013) J. Biol. Chem. 288, 28254.
    Immunohistochemistry citations
    1. Mouse brain sections (1:1000).
      Burton, S.D. et al. (2017) J. Neurosci. 37, 1117.
    2. Mouse subventricular zone sections.
      Redolfi, N. et al. (2016) Hum. Mol. Genet. 25, 5198.
    3. Mouse brain sections.
      Ghafari, M. et al. (2017) Brain Struct. Funct. 222, 549.
    4. Mouse brain sections (1:3000).
      Du, Z. et al. (2016) Neuroscience 329, 363.
    5. Rat brain sections (1:100).
      Arellano, R.O. et al. (2016) Mol. Pharmacol. 89, 63.
    6. Mouse brain sections (1:500).
      Seo, S. and Leitch, B. (2015) Neuroscience 306, 28.
    7. Mouse brain sections (1:500).
      Seo, S. and Leitch, B. (2014) Epilepsia 55, 224.
    8. 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.
    3. Rat adrenal medullary cells (1:100).
      Matsuoka, H. et al. (2008) J. Physiol. 586, 4825.
    More product citations
    1. de Luca, E. et al. (2017) Neuron 95, 63.
    2. Pennacchietti, F. et al. (2017) J. Neurosci. 37, 1747.
    3. Sengupta, J.N. et al. (2013) Pain 154, 59.
    4. Yu, X. and Hou, Z.H. (2013) J. Biol. Chem. 288, 2501.
    5. Grady, M.R. et al. (2006) J. Neurosci. 26, 2841.
    6. Mangan, P.S. et al. (2005) Mol. Pharmacol. 67, 775.
    7. Sun, C. et al. (2004) Brain Res. 1029, 207.
    8. Yazulla, S. et al. (2001) J. Neurocytology. 30, 551.
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