Synaptic Systems - Synaptotagmin1 sdAb

Synaptotagmin1 sdAb cytoplasmic tail - FluoTag-X2 - N2302-AF647-L

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

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Camelid single domain antibodies (sdAbs) consist only of one antigen binding site of an Alpaca heavy chain antibody. With only ~15 kDa, these Tags are about 10-times smaller than conventional IgG antibody molecules.

Camelid single domain antibody

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.

Unlabeled variants and several modifications of sdAbs like biotin, fluorophore or DBCO conjugation are available.

In FluoTag®-X2 two fluorophore molecules are site-specifically coupled to each FluoTag molecule. Therefore, the reagent simultaneously targets two fluorophores to the protein of interest, which ensures up to two-fold („2X“)-brighter signals. Owing to the small size of the FluoTags, the distance between the target epitope and each fluorophore is ~ 3 nm.
In comparison to detection systems using conventional antibodies, FluoTag-X can thus improve the localization accuracy by 10-15 nm. Both features - superior brightness and precise fluorophore placement - render the FluoTag-X products excellent tools for all microscopy techniques.

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

Cat. No. N2302-AF647-L	200 µl purified antibody, lyophilized from PBS, fluorescence-labeled with Alexa 647. Albumin was added for stabilization. For reconstitution add 200 µl H₂O. 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.
Storage	Up to 3 months: –20 °C Up to 12 months: –80 °C or below Protect from light!
Applications	ICC: 1 : 500 gallery IHC: 1 : 500
Label	Alexa 647, two fluorophores coupled to one FluoTag
Clone	7D5
Subtype	single domain
Immunogen	Recombinant protein corresponding to AA 80 to 421 from rat Synaptotagmin1 (UniProt Id: P21707)
Reactivity	Reacts with: mouse (P46096), rat (P21707). Other species not tested yet.

Synaptotagmin1 sdAb cytoplasmic tail - FluoTag-X2 - N2302-AF647-L

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