Tyrosine Hydroxylase, phosphorylated (Ser40) (TH, TYH, Tyrosine 3 Hydroxylase, Tyrosine 3 Monooxygenase)

Tyrosine hydroxylase (TH, tyrosine 3-monooxygenase) is the eukaryotic enzyme that catalyzes the first, rate-limiting step in the biosynthesis of dopamine and other catecholamines from tyrosine.1 TH requires O2 and ferrous iron for the hydroxylation reaction it catalyzes. In mammals, TH is encoded by a single gene that is required for embryonic development and survival;2 full-length cDNA for this enzyme has been obtained for rat, mouse, bovine, Drosophila, macaque monkey, and human species.3 In all vertebrates, TH activity is closely regulated at many levels of expression, including gene transcription, RNA processing, stability of mRNA, feedback inhibition on enzyme, and protein kinase activation.2 The active tyrosine hydroxylase enzyme is a tetramer, with its active site located within its C-terminal region, where the physiological co-substrate binds. The N-terminus acts as a regulatory domain, and contains four serine residues, including Ser 19, Ser 31, and Ser40, that can be phosphorylated by protein kinase A (PKA), CaM kinase II (CaMKII), and protein kinase C (PKC).3 Ser40 is the major TH phosphorylation site of PKA and CaMKII in vertebrates.2,3 Phosporylation of TH at Ser40, and thus the synthesis of dopamine, has been observed to be influenced by many factors, including potassium ion concentration,4 activation of adenylyl cyclase,5 NMDA receptor activity,5 ERK1/2 activity,6,7 and the binding of 14-3-3 proteins.8\n\nApplications: Suitable for use in ELISA, Western Blot, Immunoprecipitation, Immunohistochemistry. Other applications not tested.\n\nRecommended Dilution: \nELISA: 1: 1000 \nWestern Blot: 1: 1000 \nImmunohistochemistry: 1: 1000 \nImmunoprecipitation: 1: 1000\n\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability: May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at-20 degrees C or colder. Aliquots are stable for at least 12 months at-20 degrees 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.

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SPECIFICATIONS

Catalog Number

T9237-10H

Size

100ul

Applications

ELISA, IHC, IP, WB

Hosts

Rabbit

Reactivities

Rat

Form

Supplied as a liquid in PBS, 10% glycerol, 10mM HEPES, pH 7.2, 20 mM Tris-HCl, pH 7.6, 1mg/ml BSA, 0.1% PVP-40, 0.05% Tween20, 0.1% sodium azide.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Nagatsu T, et al. J Biol Chem 239:2910-2917, 1964.\n2. Vie A, et al. J Biol Chem 274(24):16788-16795, 1999.\n3. Alterio J, et al. J Biol Chem 273(17):10196-10201, 1998.\n4. Haycock JW, et al. J Neurochem 71(4):1670-1675, 1998.\n5. Lindgren N, et al. J Neurochem 74:2470-2477, 2000.\n6. Lindgren N, et al. Eur J Neurosci 15(4):769-773, 2002.\n7. Salvatore MF, et al. J Neurochem 79(2):349-360, 2001.\n8. Kleppe R, et al. J Neurochem 77(4):1097-1107, 2001.

Additional Info

Speci?c for the ~60kD rat tyrosine hydroxylase protein, phosphorylated at Ser40 and does not react with the non-phosphorylated form of the TH protein. Reactivity has been con?rmed in rat PC-12 cell lysates stimulated with okadaic acid. Based on amino acid sequence homology, this antibody is also expected to react with human and mouse.