Four different antipeptide antibodies were generated in chickens against sequences shared between the mouse (NP_033332) and human (NP_005630) gene products. Antibodies were affinity-purified and mixed in equal concentrations. These antibodies have been validated with human, mouse and rat tissues.
Immunohistochemical staining of SynTag (GREEN, Aves Labs, 1:1000 dilution) and Vimentin (RED, Rockland, 1:500 dilution) in a cryostat section through the basal plate of an e16 mouse brain.
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.
ICC, IHC, WB
SYT1 SVP65 SYT
Human, Mouse, Rat
Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.
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 concentra-tions 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.
These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.