Anti-Tyrosine Hydroxylase (TYH) Antibody

Human TYH (EC 1.14.16.2) is a 58,523 dalton protein (528 amino acids) responsible for the enzymatic conversion of L-tyrosine to L-DOPA (dihydroxyphenylalanine) . This enzyme is expressed in all catecholaminergic neurons of the CNS and PNS. In the CNS, TYH-positive neurons can be found within the substantia nigra, ventral tegmental area, locus ceruleus, and hypothalamus. In the PNS, TYH-positive neurons can be found within the sympathetic chain, pre-vertebral ganglia and the adrenal medulla.

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 two synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the Tyrosine Hydroxylase gene product, but were shared between the human (P07101, NCBI) and mouse (P24529, 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 affinity purified using a peptide column, and the concentrations of the eluates adjusted to 200 μg/mL. Finally, equal volumes of both affinity-purified anti-peptide antibody preparations were mixed, and the preparation was filter-sterilized.

Both of the antibodies were analyzed by immunohistochemistry (at a concentration of 3 μg/mL) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.