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Synaptotagmin1 antibody luminal domain - 105 311AT647N

Synaptotagmin1 is a Ca2+-sensor on synaptic vesicles that triggers neurotransmitter release
Mouse monoclonal purified IgG
Cat. No.: 105 311AT647N
Amount: 100 µg
Price: $465.00
Cat. No. 105 311AT647N 100 µg purified IgG, lyophilized, fluorescence-labeled with ATTO® 647N.

For many of the fluorescence labeled antibodies conjugated to ATTO® dyes from ATTO-TEC the established fluorescence detection systems can be used.

ATTO® 488: λex 500 nm / λem 520 nm
ATTO® 550: λex 554 nm / λem 576 nm
ATTO® 565: λex 564 nm / λem 590 nm
ATTO® 594: λex 603 nm / λem 626 nm
ATTO® 647N: λex 646 nm / λem 664 nm
 

ATTO®488, ATTO®565, ATTO®647N and ATTO®594 dyes are suitable for Stimulated Emission Depletion (STED) microscopy which allows higher resolution imaging compared to confocal laser scanning microscopy.
ATTO®488, ATTO®565 and ATTO®594 are also recommended for PALM and dSTORM high resolution microscopy.

This product or portions thereof is manufactured under license from ATTO-TEC GmbH.
ATTO is a trademarks of ATTO-TEC GmbH, Siegen/Germany.
Purchase of reagents related to the AbberiorStar 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 ATTO-TEC GmbH.

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

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 ATTO 647N
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.

Data sheet 105_311at647n.pdf

References for Synaptotagmin1 - 105 311AT647N

Presynaptic activity and protein turnover are correlated at the single-synapse level.
Jähne S, Mikulasch F, Heuer HGH, Truckenbrodt S, Agüi-Gonzalez P, Grewe K, Vogts A, Rizzoli SO, Priesemann V
Cell reports (2021) 3411: 108841. 105 311AT647N ICC; tested species: rat
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 311AT647N ICC, UPTAKE; tested species: rat
Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins.
Wilhelm BG, Mandad S, Truckenbrodt S, Kröhnert K, Schäfer C, Rammner B, Koo SJ, Claßen GA, Krauss M, Haucke V, Urlaub H, et al.
Science (New York, N.Y.) (2014) 3446187: 1023-8. 105 311AT647N ICC; 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 311AT647N ICC; tested species: rat
Limited intermixing of synaptic vesicle components upon vesicle recycling.
Opazo F, Punge A, Bückers J, Hoopmann P, Kastrup L, Hell SW, Rizzoli SO
Traffic (Copenhagen, Denmark) (2010) 116: 800-12. 105 311AT647N ICC
Neurofilament Levels in Dendritic Spines Associate with Synaptic Status.
Gürth CM, do Rego Barros Fernandes Lima MA, Macarrón Palacios V, Cereceda Delgado AR, Hubrich J, D'Este E
Cells (2023) 126: . 105 311AT647N UPTAKE; tested species: rat
Synaptic activity and strength are reflected by changes in the post-synaptic secretory pathway.
Gürth CM, Dankovich TM, Rizzoli SO, D'Este E
Scientific reports (2020) 101: 20576. 105 311AT647N UPTAKE; tested species: rat
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 311AT647N UPTAKE; tested species: rat
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 311AT647N ICC, UPTAKE; tested species: rat
STED nanoscopy reveals the ubiquity of subcortical cytoskeleton periodicity in living neurons.
D'Este E, Kamin D, Göttfert F, El-Hady A, Hell SW
Cell reports (2015) 108: 1246-51. 105 311AT647N UPTAKE; tested species: rat
The same synaptic vesicles drive active and spontaneous release.
Wilhelm BG, Groemer TW, Rizzoli SO
Nature neuroscience (2010) 1312: 1454-6. 105 311AT647N UPTAKE
Cat. No.: 105 311AT647N
Amount: 100 µg
Price: $465.00
Presynaptic activity and protein turnover are correlated at the single-synapse level.
Jähne S, Mikulasch F, Heuer HGH, Truckenbrodt S, Agüi-Gonzalez P, Grewe K, Vogts A, Rizzoli SO, Priesemann V
Cell reports (2021) 3411: 108841. 105 311AT647N ICC; tested species: rat
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 311AT647N ICC, UPTAKE; tested species: rat
Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins.
Wilhelm BG, Mandad S, Truckenbrodt S, Kröhnert K, Schäfer C, Rammner B, Koo SJ, Claßen GA, Krauss M, Haucke V, Urlaub H, et al.
Science (New York, N.Y.) (2014) 3446187: 1023-8. 105 311AT647N ICC; 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 311AT647N ICC; tested species: rat
Limited intermixing of synaptic vesicle components upon vesicle recycling.
Opazo F, Punge A, Bückers J, Hoopmann P, Kastrup L, Hell SW, Rizzoli SO
Traffic (Copenhagen, Denmark) (2010) 116: 800-12. 105 311AT647N ICC
Neurofilament Levels in Dendritic Spines Associate with Synaptic Status.
Gürth CM, do Rego Barros Fernandes Lima MA, Macarrón Palacios V, Cereceda Delgado AR, Hubrich J, D'Este E
Cells (2023) 126: . 105 311AT647N UPTAKE; tested species: rat
Synaptic activity and strength are reflected by changes in the post-synaptic secretory pathway.
Gürth CM, Dankovich TM, Rizzoli SO, D'Este E
Scientific reports (2020) 101: 20576. 105 311AT647N UPTAKE; tested species: rat
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 311AT647N UPTAKE; tested species: rat
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 311AT647N ICC, UPTAKE; tested species: rat
STED nanoscopy reveals the ubiquity of subcortical cytoskeleton periodicity in living neurons.
D'Este E, Kamin D, Göttfert F, El-Hady A, Hell SW
Cell reports (2015) 108: 1246-51. 105 311AT647N UPTAKE; tested species: rat
The same synaptic vesicles drive active and spontaneous release.
Wilhelm BG, Groemer TW, Rizzoli SO
Nature neuroscience (2010) 1312: 1454-6. 105 311AT647N UPTAKE
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.