Synaptotagmin1 antibody luminal domain - 105 311AF

Synaptotagmin1 is a Ca²⁺-sensor on synaptic vesicles that triggers neurotransmitter release

pH-sensitive probe fluorescent only under acidic conditions

Mouse monoclonal purified IgG

Cat. No.: 105 311AF

Amount: 100 µg

Price: $465.00

Cat. No. 105 311AF	100 µg purified IgG, lyophilized, 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 100 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 : 500 (see remarks) gallery IHC: not tested yet IHC-P: not tested yet EM: N/A ELISA: N/A FACS: 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. Electron microscopy (EM) is a microscopy technique that detects the scatter of electrons through thin tissue sections. In Immuno-EM the antigen is usually revealed by colloidal gold conjugated secondary antibodies linking electron dense structures to antigen-bound primary antibodies. Enzyme-linked immunosorbent assay (ELISA); a frequently employed method to quantify target-molecules in solution. The detection of some proteins requires special solubilization steps. For further information, please refer to the „Remarks“ section. Fluorescence-activated cell sorting (FACS); usually cells are fluorescently labeled with antibodies against cell-surface antigens followed by cell-sorter analysis.
Label	AcidiFluor Orange
Clone	604.2
Subtype	IgG1
Immunogen	Synthetic peptide corresponding to residues near the amino terminus of rat Synaptotagmin1 (UniProt Id: P21707)
Reactivity	Reacts with: rat (P21707). No signal: mouse (P46096), zebrafish. Other species not tested yet.
Remarks	ICC: This antibody is intended to be used for direct labeling of recycling synapses in primary neuronal cultures. The pH sensitive dye regaines its fluorescence after the reacidification of the synaptic vesicle lumen.
Data sheet	105_311af.pdf

Cat. No.: 105 311AF

Amount: 100 µg

Price: $465.00

ICC

Background

Synaptotagmin1, also known as p65, is an integral membrane glycoprotein of neuronal synaptic vesicles and secretory granules of neuroendocrine cells that is widely (but not ubiquitously) expressed in the central and peripheral nervous system. It has a variable N-terminal domain that is exposed to the lumen of the vesicle and a conserved cytoplasmic tail that contains two Ca²⁺-binding C2-domains.
Ca²⁺-binding to synaptotagmin triggers exocytosis of synaptic vesicles, thus linking Ca²⁺-influx during depolarization to neurotransmitter release.
Lumenal antibodies were used in living neurons to label synaptic vesicles from the outside via endocytotic uptake.

1 ) Geppert M et al., Annu. Rev. Neurosci. (1998)
2 ) Jahn R et al., Annu. Rev. Neurosci. (1994)
3 ) Südhof TC et al., Nature (1995)
4 ) Geppert M et al., Cell (1994)
5 ) Brose N et al., Science (1992)
6 ) Perin MS et al., Nature (1990)

Protocols

ICC

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