Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody

γ-Aminobutyric acid receptor type A subunit α1, GABRA1
    Cat #: AGA-001-AG
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    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.
    Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.
    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 -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis (unlabeled antibody, #AGA-001), and immunohistochemistry (labeled antibody).
    Applications: ic, ih, lci
    Immunohistochemistry
    Immuno-colocalization of CaV1.2 and GABA(A) α1 Receptor in rat and mouse hippocampus
    Immunohistochemical staining of mouse and rat hippocampal dentate gyrus using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG) in the same section. Both CaV1.2 (red) and GABRA1 (green) are detected in neuron-shaped cells (arrows). Staining suggests partial colocalization between CaV1.2 and GABRA1 in a sub-population of rat dentate gyrus neurons. A. Mouse hippocampus. B. Rat hippocampus.
    Expression of GABRA1 in rat brain
    Immunohistochemical staining of rat dentate gyrus using Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG) (green). Staining reveals hilar neurons (arrows) and fibers coursing through the granule layer (G). DAPI is used as the counterstain (blue).
    Immunocytochemistry
    Rat primary hippocampal neurons (Sarto-Jackson, I. et al. (2012) J. Biol. Chem. 287, 14201.).
    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)-ATTO-488 Antibody (#AGA-001-AG), (1:25), (green). B. Live view of the cell. C. Merge image of A and B.
    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)-ATTO-488 Antibody
    Immuno-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)-ATTO-488 Antibody (#AGA-001-AG). A. CaV1.2 (red) is detected mostly in Purkinje cells (arrow). B. In the same section, GABRA1 (green) is observed in the granule layer. C. Merge of the two images suggests some colocalization between CaV1.2 and GABRA1 in the rat granule layer, but only CaV1.2 appears in Purkinje cells.

    Last update: 07/11/2018

    Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001) is a highly specific antibody directed against an extracellular epitope of the rat protein. The antibody can be used in western blot, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize GABRA1 from human, rat, and mouse samples.

    Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG) is directly labeled with an ATTO-488 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-488 label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody has been tested in immunohistochemistry and live cell imaging applications and is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use