Activating sheep antibodies to tyrosine hydroxylase
Antibodies to tyrosine hydroxylase (TH; EC 188.8.131.52) have been extremely useful in establishing the neuronal and cellular localization of TH (Pickel et al., 1975a,b) and, recently, in demonstrating that in situ phosphorylation of TH is important in regulating the catalytic activity of TH (Waymire et al., 1979). While in the process of raising antibodies to TH, we observed an interesting property of immunoglobulins (Ig) from a sheep that was injected with TH purified from rat pheochromocytoma. Mixture of a partially purified TH and sheep Ig (0–45% ammonium sulfate) resulted in a dramatic elevation of TH activity. The following experiments were performed to establish whether this effect resulted from a specific interaction of TH and antibodies directed against TH. The results indicate that this crude Ig fraction contains anti-TH antibodies, that the majority of anti-TH activity is associated with activation of TH, and that the anti-TH antibodies are not uniformly distributed across IgG subclasses.
KeywordsTyrosine Hydroxylase Dramatic Elevation Couple Decarboxylation Peroxidase Label Antibody Method Tyrosine Hydroxylase Protein
Unable to display preview. Download preview PDF.
- Cinader, B. (1977). Meth. Immunol. Immunochem. 4, 313–375.Google Scholar
- Ey, P.L., Prowse, S.J. and Jenkin, C.R. (1978). Isolation of pure IgGl, IgG2a, IgG2b immunoglobulins from mouse serum usin protein A-Sepharose. Immunochem. 15, 429–436.Google Scholar
- Kelly,. P.T. and Luttges, M.W. (1975). Electrophoretic separation of nervous system proteins on exponential gradient polyacrylamide gels. J. Neurochem. 24, 1077–1079.Google Scholar
- Lloyd, T. and Kaufman, S. (1973). Production of antibodies to bovine adrenal tyrosine hydroxylase. Molec. Pharmacol. 9, 438–444.Google Scholar
- Markey, K.A., Kondo, S., Shenkman, L. and Goldstein, M. (1980). Purification and characterization of tyrosine hydroxylase from a clonal pheochromocytoma cell line. Molec. Pharmacol. 17, 79–85.Google Scholar
- Okada, Y., Ikenaka, T., Yagura, T. and Yamamura, Y. (1963). Immunological heterogeneity of rabbit antibody fragments against Taka-amylase A. J. Biochem. (Tokyo) 54, 101–102.Google Scholar
- Pollock, M.R. (1964). Stimulating and inhibiting antibodies for bacterial penicillinase. Immunol. 7, 707–723.Google Scholar
- Richmond, M.H. (1963). Purification andproperties of the exopenicillinase from Staphylococcus aureus. Biochem. J. 88, 452–459.Google Scholar
- Ross, M.E., Joh, T.H. and Reis, D.J. (1981) Monoclonal antibodies to tyrosine hydroxylase: Production and characterization. Brain Res. 207, in press.Google Scholar
- Stephens, J.K., Masserano, J.M., Vulliet, P.R., Weiner, N. and Nakame, P.K. (1981). Immunocytochemical localization of tyrosine hydroxylase in rat adrenal medulla by the peroxidase labelled antibody method. Brain Res. 207, in press.Google Scholar