Anti-Beta-Tubulin 3 (TUJ) Antibody

Human Beta-Tubulin 3 is a 50,432 dalton structural protein (450 amino acid) expressed in neurons of the PNS and CNS. It contributes to microtubule stability in neuronal cell bodies and axons, and plays a role in axonal transport.

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 three synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the β-Tubulin 3 gene product, but are shared between the human (NP_AAL28094, NCBI) and rat (AAM28438, NCBI) protein sequences. After repeated injections, 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 300 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.