Mature rat hippocampal neurons (14 DIV) were incubated live at 37°C & 5% CO₂ in full Neurobasal medium with 100 nM sdAb anti-Syt1 Luminal Domain Halo fusion (Cat. No. N4841, dilution 1:50) and 100 nM Halo ligand SiR for 30 min. After washing and PFA fixation, cells were counterstained with FluoTag®-X2 anti-Syt1 Cytosolic Domain (A51) directly conjugated to Atto488 (Cat. No. N4302-AT488-L, dilution 1:500). Images were acquired using a 60× oil-immersion objective on an epifluorescence microscope. The sdAb anti-Syt1 Luminal Domain Halo fusion labels only active/recycling synapses (A, magenta), whereas FluoTag®-X2 anti-Syt1 Cytosolic Domain (A51) labels the total Syt1 pool (B, cyan). Arrowheads indicate expected colocalization at some synapses. DAPI is displayed in yellow on the merged right panel (C). Scale bar: 20 µm.

sdAb anti-Syt1 Luminal Domain Halo fusion

Name: sdAb anti-Syt1 Luminal Domain Halo fusion
SKU: N4841
Availability: InStock

Cat No: N4841 Category: Primary sdAbs

290,00 €

A Halo-tagged version of a single-domain antibody recognizing the luminal/extracellular domain of Synaptotagmin 1 (Syt1). This reagent is intended exclusively for use in living neurons to label active synapses that recycle Syt1 following synaptic vesicle fusion.

sdAb anti-Syt1 Luminal Domain Halo fusion combines a single-domain antibody (sdAb) targeting the luminal/extracellular domain of Synaptotagmin 1 (Syt1) with a dehalogenase-based technology. The HaloTag^®⁽¹⁾sequence is fused to the C-terminus of the sdAb anti-Syt1 Luminal Domain. This fusion enables highly flexible and selective labeling using a variety of Halo ligands. Cell-permeable or cell-impermeable Halo ligands can be applied to label different pools of synaptic vesicles or surface Synaptotagmin 1 in live-cell systems.

Syt1 belongs to the synaptotagmin protein family, which in mammals comprises 17 single-pass transmembrane proteins containing two intracellular calcium-binding domains. Synaptotagmin 1 is enriched at presynaptic boutons on axon terminals, where it is associated with synaptic vesicles, and implicated in triggering vesicle fusion and neurotransmitter release upon a local increase in calcium concentration.

Our sdAb anti-Syt1 Luminal Domain binds specifically to the mouse and rat Syt1 luminal/extracellular epitope KEDAFSKLKQKF, and the human epitope KEDAFSKLKEKF (other species not tested).

Important Note: The reagent is intended exclusively for use in live-cell systems to label active synapses. Aldehyde-based fixatives (including paraformaldehyde, glutaraldehyde, glyoxal, or related compounds) modify lysines within the target epitope, thereby significantly impairing Syt1 recognition.

Comprehensive characterization data for this sdAb anti-Syt1 Luminal Domain have been published here, where it is referred to as NbLumSyt1.

sdAb anti-Syt1 Luminal Domain (clone #1F12) binds to the extracellular/luminal domain of Syt1 in non-fixed samples. We also offer reagents based on sdAbs anti-Syt1 Cytosolic Domain (clones #A51 and #7D5), including labeled sdAbs, FluoTags^®-X2, and recombinant heavy-chain antibodies (rHcAbs). These reagents specifically recognize the cytosolic portion of Syt1 in fixed and non-fixed samples, thereby enabling detection of the total Syt1 pool. View the reagents targeting the Syt1 cytosolic domain here.

⁽¹⁾ HaloTag^® is a registered trademark of Promega Corporation.

MSDS