Anti-VGLUT2 Antibody

Vesicular Glutamate Transporter 2, DNPI, SLC17A6
    Cat #: AGC-036
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)EDGKPLEVPEKK, corresponding to amino acid residues 45-56 of rat VGLUT2 (Accession Q9JI12). Cytoplasmic, N-terminus.
    Accession (Uniprot) Number Q9JI12
    Gene ID 84487
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human - 11/12 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 -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ih, wb
    May also work in: ic, ifc, ip
    Western blot
    Western blot analysis of rat brain membranes (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and human SH-SY5Y neuroblastoma cell lysate (lanes 3 and 6):
    1-3. Anti-VGLUT2 Antibody (#AGC-036), (1:400).
    4-6. Anti-VGLUT2 Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of VGLUT2 in rat neocortex
    Immunohistochemical staining of rat frozen brain sections using Anti-VGLUT2 Antibody (#AGC-036), (1:400). VGLUT2 (red) is detected in pyramidal neurons (arrow). The same section was stained with DAPI (blue).
    Immuno-colocalization of VGLUT2 and P2X7 Receptor in rat spinal cord
    Immunohistochemical staining of perfusion-fixed frozen rat spinal cord sections using Anti-VGLUT2 Antibody (#AGC-036), (1:600) and Anti-P2X7 Receptor-ATTO-550 Antibody (#APR-004-AO), (1:100). A. Vesicular Glutamate Transporter 2 labeling followed by goat-anti-rabbit-Alexa-488 (green). B. The same section labeled for P2X7 Receptor (orange). C.  Merge of A and B demonstrates partial co-localization of VGLUT2 and P2X7 Receptor in dorsal horn and in lateral column (L. Col., arrow). Cell nuclei were stained with DAPI (blue).
    References
    1. Gong, J. et al. (2006) Brain Res. 1082, 73.
    2. Fremeau, R.T. et al. (2004) Science 304, 1815.
    3. Takamori, S. et al. (2000) Nature 407, 189.
    4. Maycox, P.R. et al. (1988) J. Biol. Chem. 263, 15423.
    5. Gras, C. et al. (2002) J. Neurosci. 22, 5442.
    6. Moechars, D. et al (2006) J. Neurosci. 26, 12055.
    Scientific background

    Central nervous system neurons have traditionally been thought to express exclusively membrane transporters and/or vesicular transporters for their transmitter. Three vesicular glutamate transporters (VGLUTs) have been cloned: BNPI/VGLUT1 (a brain-specific sodium dependent inorganic phosphate (Pi) transporter), and its homologs DNPI/VGLUT2 (differentiation-associated sodium-dependent Pi transporter) and VGLUT31. These transporters mediate glutamate uptake inside presynaptic vesicles and are anatomical and functional markers of glutamatergic excitatory transmission2.

    BNPI/DNPI encodes a membrane protein with 6-8 putative transmembrane domains which exhibits weak similarities to a kidney Na+-dependent inorganic phosphate co-transporter3. The transporters use a membrane potential gradient set by the vesicular H+-ATPase for glutamate uptake4. VGLUT1-3 are very similar in structure and function, but are used by different neuronal populations. VGLUT1 and VGLUT2 are expressed by the cortical and subcortical neurons respectively. VGLUT3 is expressed by non-glutamatergic neurons5. VGLUT2 is expressed in the thalamus, brainstem, and deep cerebellar nuclei2. A Recent study has shown that  targeted deletion of VGLUT2 in mice causes perinatal lethality and a 95% reduction in evoked glutamatergic responses in thalamic neurons, although hippocampal synapses function normally. Behavioral analysis of heterozygous VGLUT2 mice showed discrete behavioral phenotypes that suggest a deficit in thalamic processing6.

    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 VGLUT2 and P2X7 Receptor in rat spinal cordImmunohistochemical staining of perfusion-fixed frozen rat spinal cord sections using Anti-VGLUT2 Antibody (#AGC-036), (1:600) and Anti-P2X7 Receptor-ATTO-550 Antibody (#APR-004-AO), (1:100). A. VGLUT2 labeling followed by goat-anti-rabbit-Alexa-488 (green). B. The same section labeled for P2X7 Receptor (orange). C.  Merge of A and B demonstrates partial co-localization of Vesicular Glutamate Transporter 2 and P2X7 Receptor in the ventral horn (arrow). Cell nuclei were stained with DAPI (blue).

    Last update: 21/11/2018

    Anti-VGLUT2 Antibody (#AGC-036) is a highly specific antibody directed against an epitope of the rat Vesicular Glutamate Transporter 2. The antibody can be used in western blot analysis and immunohistochemistry applications. It has been designed to recognize VGLUT2 from rat, mouse and human samples.

    For research purposes only, not for human use