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Synaptotagmin1 antibody luminal domain - 105 103CpH

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

pH-sensitive probe fluorescent only under acidic conditions

Rabbit polyclonal purified antibody
Cat. No.: 105 103CpH
Amount: 50 µg
Price: $485.00
Cat. No. 105 103CpH 50 µg specific antibody, lyophilized. Affinity purified with the immunogen, fluorescence-labeled with CypHer5E.

CypHer5E® is a pH-sensitive dye, fluorescent only after antibody internalization at the acidic pH of the lumen of the synaptic vesicle. CypHer5E® can be detected with the filter sets for Cy5®.

CypHer5E®: λex 647 nm / λem 667 nm

This product or portions thereof is manufactured under license from Carnegie Mellon University under U.S. Patent Application Number US 2006-0051758 and related patents. Cy, CyDye, and CypHer are trademarks of GE-Healthcare Ltd. Purchase of reagents related to the CypHer 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 GE-Healthcare Bio-Science Corp., 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.

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 : 500 (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 CypHer5E
Immunogen Synthetic peptide corresponding to AA 1 to 8 from mouse Synaptotagmin1 (UniProt Id: P46096)
Reactivity Reacts with: rat (P21707), mouse (P46096).
Other species not tested yet.

For unknown reasons antibodies raised against the luminal N-terminus of Synaptotagmin 1 show a strong preference for the rat protein.
Matching control protein/peptide 105-10P
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_103cph.pdf
Cat. No.: 105 103CpH
Amount: 50 µg
Price: $485.00
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.