Anti-Vesicular GABA Transporter Antibody

VGAT, VIAAT, SLC32A1
    Cat #: AGT-005
  • 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.
    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 1 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, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat brain membranes (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and human U-87 MG glyoblastoma lysates (lanes 3 and 6):
    1-3. Anti-Vesicular GABA Transporter Antibody (#AGT-005), (1:200).
    4-6. Anti-Vesicular GABA Transporter Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of Vesicular GABA Transporter in rat cerebellum
    Immunohistochemical staining of rat frozen cerebellum sections using Anti-Vesicular GABA Transporter Antibody (#AGT-005), (1:100). VGAT staining is shown in red and dapi (blue) was used as a general cellular marker. VGAT appears in the cerebellar pinceau (arrows) and in the soma of Purkinje cells (triangles).
    Immunocytochemistry
    Colocalization of GluR1 and Vesicular GABA Transporter in human U-87 MG cells
    Immunocytochemical staining of human glioblastoma U-87 MG. Extracellular staining of live intact cells with Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009), (1:25), followed by goat anti-guinea pig-AlexaFluor-488 secondary antibody (green). Cells were subsequently fixed, permeabilized and labeled with Anti-Vesicular GABA Transporter Antibody (#AGT-005), (1:200), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). Representative merged images of the double labeled cells are shown in A and B.
    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. Chaudry, F. 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
    Last update: 29/11/2018

    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.

    For research purposes only, not for human use