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VGAT antibody luminal domain - 131 103C2 K.O.

VGAT or VIAAT is the GABA and glycine transporter in the membrane of synaptic vesicles
Rabbit polyclonal purified antibody
Cat. No.: 131 103C2
Amount: 50 µg
Price: $780.00
Cat. No. 131 103C2 50 µg specific antibody, lyophilized. Affinity purified with the immunogen, fluorescence-labeled with Cyanine 2.

Fluorescence labeled antibodies conjugated to (Sulfo-)Cyanine dyes are well suited for standard epi-fluorescence setups and confocal microscopy.

Cyanine 2: λex 492 nm / λem 508 nm
Sulfo-Cyanine 3: λex 554 nm / λem 568 nm
Sulfo-Cyanine 5: λex 649 nm / λem 667 nm

(Sulfo-)Cyanine 5 dyes are well suited for STORM high resolution microscopy.
Sulfo-Cyanines are highly water soluble and allow a higher labeling degree and consequently brighter conjugates.

Albumin was added for stabilization. For reconstitution add 50 µl H2O to get a 1mg/ml solution in PBS. Either add 1:1 (v/v) glycerol, then aliquot and store at -20°C until use, or store aliquots at -80°C without additives.
Reconstitute immediately upon receipt! Avoid bright light when working with the antibody to minimize photo bleeching of the fluorescent dye.
Applications
 
WB: N/A
IP: N/A
ICC: 1 : 100 up to 1 : 200 (see remarks) gallery  
IHC: yes
IHC-P: not tested yet

Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section.

Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section.

Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section.

Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section.

Immunohistochemistry (IHC-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.

Label Cyanine 2
Immunogen Synthetic peptide corresponding to residues near the carboxy terminus of rat VGAT (UniProt Id: O35458)
Reactivity Reacts with: human (Q9H598), rat (O35458), mouse (O35633).
Other species not tested yet.
Specificity K.O. validated
Remarks

ICC: This antibody is intended for labeling of recycling synaptic vesicles at inhibitory nerve terminals by adding to living neurons. Further details see Martens et al. 2008.

Data sheet 131_103c2.pdf

References for VGAT - 131 103C2

Clathrin-independent endocytic retrieval of SV proteins mediated by the clathrin adaptor AP-2 at mammalian central synapses.
López-Hernández T, Takenaka KI, Mori Y, Kongpracha P, Nagamori S, Haucke V, Takamori S
eLife (2022) 11: . 131 103C2 ICC; tested species: mouse
A ratiometric ER calcium sensor for quantitative comparisons across cell types and subcellular regions.
Farrell RJ, Bredvik KG, Hoppa MB, Hennigan ST, Brown TA, Ryan TA
bioRxiv : the preprint server for biology (2024) : . 131 103C2 UPTAKE; tested species: rat
Key physiological parameters dictate triggering of activity-dependent bulk endocytosis in hippocampal synapses.
Wenzel EM, Morton A, Ebert K, Welzel O, Kornhuber J, Cousin MA, Groemer TW
PloS one (2012) 76: e38188. 131 103C2 UPTAKE
Stabilization of GABA(A) receptors at endocytic zones is mediated by an AP2 binding motif within the GABA(A) receptor β3 subunit.
Smith KR, Muir J, Rao Y, Browarski M, Gruenig MC, Sheehan DF, Haucke V, Kittler JT
The Journal of neuroscience : the official journal of the Society for Neuroscience (2012) 327: 2485-98. 131 103C2 UPTAKE
Cat. No.: 131 103C2
Amount: 50 µg
Price: $780.00
Clathrin-independent endocytic retrieval of SV proteins mediated by the clathrin adaptor AP-2 at mammalian central synapses.
López-Hernández T, Takenaka KI, Mori Y, Kongpracha P, Nagamori S, Haucke V, Takamori S
eLife (2022) 11: . 131 103C2 ICC; tested species: mouse
A ratiometric ER calcium sensor for quantitative comparisons across cell types and subcellular regions.
Farrell RJ, Bredvik KG, Hoppa MB, Hennigan ST, Brown TA, Ryan TA
bioRxiv : the preprint server for biology (2024) : . 131 103C2 UPTAKE; tested species: rat
Key physiological parameters dictate triggering of activity-dependent bulk endocytosis in hippocampal synapses.
Wenzel EM, Morton A, Ebert K, Welzel O, Kornhuber J, Cousin MA, Groemer TW
PloS one (2012) 76: e38188. 131 103C2 UPTAKE
Stabilization of GABA(A) receptors at endocytic zones is mediated by an AP2 binding motif within the GABA(A) receptor β3 subunit.
Smith KR, Muir J, Rao Y, Browarski M, Gruenig MC, Sheehan DF, Haucke V, Kittler JT
The Journal of neuroscience : the official journal of the Society for Neuroscience (2012) 327: 2485-98. 131 103C2 UPTAKE
Background
The vesicular GABA transporter VGAT is responsible for uptake and storage of GABA and glycine by synaptic vesicles in the central nervous system. For this reason it is frequently referred to as the vesicular inhibitory aminoacid transporter VIAAT. It is different from the plasma membrane transporters in that it is driven by a proton electrochemical gradient across the vesicle membrane. So far, only one isoform is known. VGAT is currently the best marker for inhibitory nerve terminals.