Anti-phospho-Tyrosine Hydroxylase (Ser40) (TH, TYH, Tyrosine 3 Hydroxylase, Tyrosine 3 Monooxygenase

Rat tyrosine hydroxylase (TH) is a pterin-dependent monooxygenase that catalyzes the hydroxylation of tyrosine to DOPA through the use of a non-heme iron. It is a 55 kD, 498 amino acid (aa) alpha-helical protein that runs anomalously at 62 kD in SDS-page. The molecule contains a 164 aa N-terminal regulatory region and a 334 aa C-terminal catalytic domain. Its activity is regulated at the post-transcriptional level by phosphorylation of serine and feedback by catecholamines. Four serines are known to be phosphorylated at aa positions 8, 19, 31, and 40. Different kinases contribute to different phosphorylation patterns. For example, MAPKAPK-2 and CaMKII act on S19 and S40, PKA phosphorylates only at S40, while Cdk5 phosphorylates S31. Variable site phosphorylations have variable effects. S40 phosphorylation blocks catecholamine feedback inhibition. Subsequent phosphorylations at S19 or S31 likely stabilize an otherwise unstable enzyme phosphorylated only at S40, contributing to increased enzyme activity. Phosphorylation at S8 is likely to be physiologically unimportant. Applications: Suitable for use in Western Blot, Immunofluorescence, Immunohistochemistry and ELISA. Other applications not tested. Recommended Dilution: Western Blot: 1:1000, Immunofluorescence: 1:1000 (frozen sections), Immunohistochemistry: 1:1000 (frozen sections), Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.