Tyrosine Hydroxylase (TH) is the rate-limiting enzyme in the synthesis of the catecholamines dopamine and norepinephrine. TH antibodies are useful markers for dopaminergic and noradrenergic neurons. Short-term regulation TH is accomplished by dynamic changes in the phosphorylation state of the enzyme. Four phosphorylation sites in TH have been identified (Ser8, Ser19, Ser31, and Ser40), all in the amino-terminal regulatory region. Cyclin-dependent kinase 5 (Cdk5) is involved in multiple aspects of neurotransmission in both post- and presynaptic compartments. Cdk5 has been described as the key enzyme for synthesis of dopamine and phosphorylates tyrosine hydroxylase at Ser31. Cdk5 also regulates ERK1/2-dependent phosphorylation of TH through the phosphorylation of mitogen-activated protein kinase kinase 1 (MEK1). Phospho-Ser31 TH levels have been found to increase in dopaminergic neurons of rats trained to chronically self-administer cocaine, suggesting that direct and indirect regulation of the phosphorylation state of the Ser31 site on TH by Cdk5/ERK1/2 may be involved in the neuroadaptive changes associated with chronic cocaine exposure.
Applications:
Suitable for use in Western Blot, Immunocytochemistry, Immunofluorescence and Immunohistochemistry. Other applications not tested.
Recommended Dilution:
Immunocytochemistry: 1:1000
Immunofluorescence: 1:1000
Immunohistochemistry (frozen): 1:1000
Western Blot: 1:1000
Optimal dilutions to be determined by the researcher.
Positive Control:
Okadaic stimulated PC-12 (1hr, 1uM)
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.