Anti-VGLUT2-ATTO-594 Antibody

Vesicular Glutamate Transporter 2, DNPI, SLC17A6
    Cat #: AGC-036-AR
  • 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.
    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, #AGC-036), and immunohistochemistry (labeled antibody).
    Applications: ih
    May also work in: ic
    Immunohistochemistry
    Immuno-colocalization of VGLUT2 and Neurokinin Receptor 1 in rat DRG
    Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglion (DRG) sections using Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60) and Anti-Neurokinin Receptor 1 (NK1) (extracellular)-ATTO-488 Antibody (#ATR-001-AG), (1:60). A. VGLUT2 staining (red). B. Neurokinin Receptor 1 staining (green). C. Merge of the two images demonstrates co-localization in some neuronal bodies (arrows point at examples). Cell nuclei are stained with DAPI (blue).
    Immuno-colocalization of AT2 Receptor and VGLUT2 in rat supraoptic hypothalamic nucleus
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG), (1:60) and Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60). A. AT2 Receptor staining (green). B. VGLUT2 staining (red). C. Merge of the two images shows a general lack of co-localization of AT2 and VGLUT2 in this part of the hypothalamus (arrow). Cell nuclei are 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 Parathyroid Hormone 2 Receptor and VGLUT2 in mouse hippocampus.Immunohistochemical staining of mouse hippocampal CA3 region frozen sections using Anti-Parathyroid Hormone 2 Receptor (extracellular)-ATTO-488 Antibody (#APR-052-AG), (1:60) and Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60). A. PTH2 receptor staining (green). B. VGLUT2 staining (red) in same section. C. Merge of A and B suggests extensive localization of PTH2R on glutamatergic neurons.

    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.

    Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-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-VGLUT2-ATTO-594 Antibody has been tested in immunohistochemical applications and is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use