Anti-GABA(A) ε Receptor (extracellular)-ATTO-633 Antibody

GABA(A) receptor subunit epsilon, Gabre, GABA(A) ε
    Cat #: AGA-015-FR
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    Immunogen
    Peptide (C)RLRYNDTFETLILH, corresponding to amino acid residues 131-144 of rat GABA(A) ε receptor (Accession Q9ES14). Extracellular, N-terminus.
    Accession (Uniprot) Number Q9ES14
    Gene ID Q9ES14
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human - 10/14 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-633. Maximum absorption 629 nm; maximum fluorescence 657 nm. The fluorescence is excited most efficiently in the 610 - 645 nm range. This label is analogous to the dyes Alexa 647, Alexa 633 and Cy5 and can be used for direct flow cytometry (FACS) using the He:Ne laser.
    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 -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis (unlabeled antibody,#AGA-015), and immunocytochemistry (labeled antibody).
    Applications: ic, ih, lci
    Immunohistochemistry
    Immuno-colocalization of Neurexin 1α and GABA(A) ε Receptor in mouse cerebellum
    Immunohistochemical staining of rat cerebellum frozen section using Anti-Neurexin 1α (extracellular)-ATTO-488 Antibody (#ANR-031-AG) and Anti-GABA(A) ε Receptor (extracellular)-ATTO-633 Antibody (#AGA-015-FR). A. Neurexin 1α staining (green). B. GABA(A) ε Receptor staining (purple). C. Merge of the two images suggests extensive co-localization in Purkinje cells (arrow).
    Live cell imaging / Immunocytochemistry
    Expression of GABA(A) ε Receptor in rat U-87 MG cells
    Immunocytochemical staining of intact living rat U-87 MG cells. A and B. Extracellular staining of cells using Anti-GABA(A) ε Receptor (extracellular)-ATTO-633 Antibody (#AGA-015-FR), (1:25), (purple) merged with live view of the cells.
    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. 19, 7057.
    5. Olsen, R.W. and Tobbin, A.J. (1990) FASEBS J. 4, 1469.
    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 system1.

    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 superfamily2. GABA inhibits the activity of signal-receiving neurons by interacting with the GABAA receptor on these cells3. 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 activity2.

    GABAARs are heteropentamers, in which all five subunits contribute to the pore formation. To date, eight subunit isoforms have been cloned: α, β, γ, δ, ε, π, θ, and ρ1. Six α subunit isoforms have been found to exist in mammals (α1-α6). In most cases, native GABAA receptors consist of 2α, 2β, and 1γ subunits. The α subunit is the most common and is expressed ubiquitously. It determines the affinity of GABAARs for allosteric ligands.

    Each subtype has a unique regional expression in the brain, and individual neurons often express multiple subtypes4. For example, the α4 subunit is detected in the hippocampus, cortex, olfactory bulb and in the basal forebrain5.

    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
    Last update: 22/10/2018

    Anti-GABA(A) ε Receptor (extracellular) Antibody (#AGA-015) is a highly specific antibody directed against an extracellular epitope of the rat GABA(A) ε subunit. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize GABA(A) ε receptor from human, rat and mouse samples.

    Anti-GABA(A) ε Receptor (extracellular)-ATTO-633 Antibody (#AGA-015-FR) is directly labeled with an ATTO-633 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. ATTO 633 has a maximum absorption at 629 nm and a maximum fluorescence at 657 nm. The fluorescence is excited most efficiently in the range 610 to 645 nm. This label is analogous to the well-known dyes Alexa 647, Alexa 633 and Cy5. Anti-GABA(A) ε Receptor (extracellular)-ATTO-633 Antibody has been tested in live cell imaging and immunohistochemistry applications. It is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use