Synaptic Systems - Synaptotagmin1

Cat. No.: 105 311CpH

Amount: 100 µg

Price: $465.00

Cat. No. 105 311CpH	100 µg purified IgG, lyophilized, 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^®: λ_ex 647 nm / λ_em 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 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 : 50 up to 1 : 300 (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	CypHer5E
Clone	604.2
Subtype	IgG1 (κ light chain)
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_311cph.pdf

References for Synaptotagmin1 - 105 311CpH

Transient Confinement of CaV2.1 Ca2+-Channel Splice Variants Shapes Synaptic Short-Term Plasticity.
Heck J, Parutto P, Ciuraszkiewicz A, Bikbaev A, Freund R, Mitlöhner J, Alonso M, Fejtova A, Holcman D, Heine M
Neuron (2019) : . 105 311CpH ICC; tested species: rat

Riluzole attenuates the efficacy of glutamatergic transmission by interfering with the size of the readily releasable neurotransmitter pool.
Lazarevic V, Yang Y, Ivanova D, Fejtova A, Svenningsson P
Neuropharmacology (2018) : . 105 311CpH ICC; tested species: rat

Regulated Dynamic Trafficking of Neurexins Inside and Outside of Synaptic Terminals.
Neupert C, Schneider R, Klatt O, Reissner C, Repetto D, Biermann B, Niesmann K, Missler M, Heine M
The Journal of neuroscience : the official journal of the Society for Neuroscience (2015) 3540: 13629-47. 105 311CpH ICC

Dynamic properties of the alkaline vesicle population at hippocampal synapses.
Röther M, Brauner JM, Ebert K, Welzel O, Jung J, Bauereiss A, Kornhuber J, Groemer TW
PloS one (2014) 97: e102723. 105 311CpH ICC; tested species: rat

Synapse clusters are preferentially formed by synapses with large recycling pool sizes.
Welzel O, Tischbirek CH, Jung J, Kohler EM, Svetlitchny A, Henkel AW, Kornhuber J, Groemer TW
PloS one (2010) 510: e13514. 105 311CpH ICC

α-Synuclein induced cholesterol lowering increases tonic and reduces depolarization-evoked synaptic vesicle recycling and glutamate release.
Lazarevic V, Yang Y, Paslawski W, Svenningsson P
NPJ Parkinson's disease (2022) 81: 71. 105 311CpH UPTAKE; tested species: mouse

Rho-kinase inhibition by fasudil modulates pre-synaptic vesicle dynamics.
Saal KA, Warth Pérez Arias C, Roser AE, Christoph Koch J, Bähr M, Rizzoli SO, Lingor P
Journal of neurochemistry (2020) : . 105 311CpH UPTAKE; tested species: rat

CtBP1-Mediated Membrane Fission Contributes to Effective Recycling of Synaptic Vesicles.
Ivanova D, Imig C, Camacho M, Reinhold A, Guhathakurta D, Montenegro-Venegas C, Cousin MA, Gundelfinger ED, Rosenmund C, Cooper B, Fejtova A, et al.
Cell reports (2020) 307: 2444-2459.e7. 105 311CpH UPTAKE; tested species: mouse

Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission.
Truckenbrodt S, Viplav A, Jähne S, Vogts A, Denker A, Wildhagen H, Fornasiero EF, Rizzoli SO
The EMBO journal (2018) : . 105 311CpH UPTAKE; tested species: rat

The pH probe CypHer™5E is effectively quenched by FM dyes.
Welzel O, Loy K, Tischbirek CH, Tabor A, Gmeiner P, Kornhuber J, Groemer TW
Journal of fluorescence (2013) 233: 487-94. 105 311CpH UPTAKE; tested species: rat

Blocking endocytosis enhances short-term synaptic depression under conditions of normal availability of vesicles.
Hua Y, Woehler A, Kahms M, Haucke V, Neher E, Klingauf J
Neuron (2013) 802: 343-9. 105 311CpH 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. 105 311CpH UPTAKE

Independent vesicle pools underlie different modes of release during neuronal development.
Andreae LC, Fredj NB, Burrone J
The Journal of neuroscience : the official journal of the Society for Neuroscience (2012) 325: 1867-74. 105 311CpH UPTAKE

Use-dependent inhibition of synaptic transmission by the secretion of intravesicularly accumulated antipsychotic drugs.
Tischbirek CH, Wenzel EM, Zheng F, Huth T, Amato D, Trapp S, Denker A, Welzel O, Lueke K, Svetlitchny A, Rauh M, et al.
Neuron (2012) 745: 830-44. 105 311CpH UPTAKE

A readily retrievable pool of synaptic vesicles.
Hua Y, Sinha R, Thiel CS, Schmidt R, Hüve J, Martens H, Hell SW, Egner A, Klingauf J
Nature neuroscience (2011) 147: 833-9. 105 311CpH UPTAKE

Amyloid precursor protein is trafficked and secreted via synaptic vesicles.
Groemer TW, Thiel CS, Holt M, Riedel D, Hua Y, Hüve J, Wilhelm BG, Klingauf J
PloS one (2011) 64: e18754. 105 311CpH UPTAKE

Systematic heterogeneity of fractional vesicle pool sizes and release rates of hippocampal synapses.
Welzel O, Henkel AW, Stroebel AM, Jung J, Tischbirek CH, Ebert K, Kornhuber J, Rizzoli SO, Groemer TW
Biophysical journal (2011) 1003: 593-601. 105 311CpH UPTAKE

A common origin of synaptic vesicles undergoing evoked and spontaneous fusion.
Hua Y, Sinha R, Martineau M, Kahms M, Klingauf J
Nature neuroscience (2010) 1312: 1451-3. 105 311CpH UPTAKE

Cat. No.: 105 311CpH

Amount: 100 µg

Price: $465.00

A readily retrievable pool of synaptic vesicles.
Hua Y, Sinha R, Thiel CS, Schmidt R, Hüve J, Martens H, Hell SW, Egner A, Klingauf J
Nature neuroscience (2011) 147: 833-9. 105 311CpH UPTAKE

A common origin of synaptic vesicles undergoing evoked and spontaneous fusion.
Hua Y, Sinha R, Martineau M, Kahms M, Klingauf J
Nature neuroscience (2010) 1312: 1451-3. 105 311CpH UPTAKE

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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Synaptotagmin1 antibody luminal domain - 105 311CpH

References for Synaptotagmin1 - 105 311CpH