Anti-Synaptotagmin-1 Antibody

Human Synaptotagmin-1 is a 47,442 dalton protein (422 amino acids) associated with pre-synaptic terminals of the PNS and CNS. Synaptotagmin-1 binds calcium and initiates the fusion of vesicles with the pre-synaptic membrane. This protein also is believed to be the target of proteolytic activity associated with Botulinum neurotoxin A.

Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least twelve months, provided they remain sterile. For longer term storage, aliquot and freeze to avoid freeze-thaw of the antibody.

Chickens were immunized with four synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the Synaptotagmin-1 gene product, but are shared between the human (NP_005630, NCBI) and mouse (NP_033332, NCBI) sequences. After repeated injections into the hens, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity purified using a peptide column, and the concentrations of the eluates adjusted to 400 μg/mL. Finally, equal volumes of each of the three affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.

Each of the three antibodies were analyzed by immunohistochemistry (at a dilution of 1:2000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.