VGAT antibody luminal domain - 131 103AF

VGAT or VIAAT is the GABA and glycine transporter in the membrane of synaptic vesicles

pH-sensitive probe fluorescent only under acidic conditions

Rabbit polyclonal purified antibody

Cat. No.: 131 103AF

Amount: 50 µg

Price: $780.00

Cat. No. 131 103AF	50 µg specific antibody, lyophilized. Affinity purified with the immunogen, fluorescence-labeled with AcidiFluor Orange. AcidiFluor Orange^® is a pH-sensitive dye, fluorescent only after antibody internalization at the acidic pH of the lumen of the cellular compartment e.g. the synaptic vesicle. It can be detected with established fluorescence detection systems. AcidiFluor Orange: λ_ex 570 nm / λ_em 590 nm This product or portions thereof is manufactured under license from Tokyo University and is covered by Patents Application Number US 2010068733, US 20140248654, EP2098529A1/B1, JP WO 2013035767A, JP 5636080B2, WO 2008/0599910A1, WO 2016/1579937A1 and issued patents US 8258171 and US 9784732 and related patents. Acidifluor®, and Acidifluor Orange® and related products are trademarks of Goryo Chemical. Purchase of reagents related to the Acidifluor Series technology from Synaptic Systems GmbH provides a license for non-profit and in-house research use only. Any application of above mentioned technology for commercial purpose requires a separate license from: Goryo Chemical, EAREE Bldg., SF Kita 8 Nishi 18-35-100, Chuo-Ku, Sapporo, 060-0008 Japan. 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 : 300 (see remarks) gallery IHC: not tested yet 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	AcidiFluor Orange
Immunogen	Synthetic peptide corresponding to residues near the carboxy terminus of rat VGAT (UniProt Id: O35458)
Reactivity	Reacts with: mouse (O35633), rat (O35458). Other species not tested yet.
Remarks	ICC: This antibody is intended for direct labeling of recycling synaptic vesicles at inhibitory nerve terminals in living primary neuronal cultures. The pH sensitive dye regaines its fluorescence after the reacidification of the synaptic vesicle lumen. Further details see Martens et al. 2008.
Data sheet	131_103af.pdf

Cat. No.: 131 103AF

Amount: 50 µg

Price: $780.00

ICC

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.

1 ) Chaudhry FA et al., J. Neurosci. (1998)
2 ) McIntire SL et al., Nature (1997)
3 ) Sagné C et al., FEBS Lett. (1997)
4 ) Hell JW et al., EMBO J. (1988)

Protocols

ICC

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