Anti-Vesicular GABA Transporter-ATTO-594 Antibody

VGAT, VIAAT, SLC32A1
    Cat #: AGT-005-AR
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)GEFGGHDKPKITAWE, corresponding to amino acid residues 106-120 of rat vesicular GABA transporter (Accession O35458). Cytoplasmic, N-terminus.
    Accession (Uniprot) Number O35458
    Gene ID 83612
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, human - 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-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
    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, #AGT-005), and immunohistochemistry (labeled antibody).
    Applications: ih
    May also work in: ic
    Immunohistochemistry
    Expression of Vesicular GABA Transporter in rat cerebellum
    Immunohistochemical staining of rat frozen cerebellum sections using Anti-Vesicular GABA Transporter-ATTO-594 Antibody (#AGT-005-AR), (1:100). A. VGAT staining is shown in red. B. DAPI is used as a general cellular marker (blue). C. Merge of A and B reveal that VGAT is most intensely expressed in the granule and pyramidal layer (arrow).
    References
    1. Gasnier, B. (2004) Pflugers Arch. 447, 756.
    2. McIntire, S. et al. (1997) Nature 389, 870.
    3. Bedet, C. et al. (2000) J. Neurochem. 75, 1654.
    4. Chaudhry, F.A. et al. (1998) J. Neurosci. 18, 9733.
    5. Dumoulin, A. et al. (1999) J. Cell Sci. 112, 811.
    6. Takamori, S. et al. (2000) J. Neurosci. 20, 4904.
    7. Mayerhofer, A. et al. (2001) FASEB J. 15, 1089.
    8. Chessler, S. et al. (2002) Diabetes 51, 1763.
    Scientific background

    The superfamily of amino acid transporters includes SLC32, SLC36 and SLC38 families. SLC32 has only one family member, the vesicular inhibitory amino acid transporter (VIAAT) or vesicular GABA transporter (VGAT)1.

    VGAT is responsible for the uptake of GABA and Glycine in exchange of protons, and their release from nerve terminals1.

    The transporter is organized into ten membrane spanning domains, a cytosolic N-terminus and a luminal C-terminal tail2. In many regions in the brain, where it is expressed, it is found constitutively phosphorylated, although the biological significance is currently unknown3.

    VGAT is expressed on synaptic vesicles of GABAergic and glycinergic neurons4-6. The transporter is also detected in the pituitary gland and in the pancreas7,8.

    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:

    Immuno-colocalization of Vesicular GABA Transporter and Gephyrin in rat spinal cord.Immunohistochemical staining of perfusion-fixed frozen rat spinal cord sections using Anti-Gephyrin Antibody (#AIP-005), (1:300) and Anti-Vesicular GABA Transporter-ATTO-594 Antibody (#AGT-005-AR), (1:80).  A. Gephyrin staining (green) appears in the superficial layer of the spinal dorsal horn and some cellular profiles were stained. B. Vesicular GABA Transporter staining (red) appears in superficial layer and in cell profiles. C. Merge images of A and B reveals co-localization (arrows) in superficial layers and separate expression in some cells. Cell nuclei were stained with DAPI (blue) as the counterstain.

    Last update: 27/11/2018

    Anti-Vesicular GABA Transporter Antibody (#AGT-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. It has been designed to recognize VGAT from human, mouse, and rat samples.

    Anti-Vesicular GABA Transporter-ATTO-594 Antibody (#AGT-005-AR) is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-Vesicular GABA Transporter-ATTO-594 Antibody has been tested in immunohistochemical applications and is especially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use