This antibody is specific for the ~55 kDa TPH in Western blots of human dorsal Raphe nucleus lysates. Immunolabeling has been demonstrated in rat brain lysates. Based upon the relatively high degree of homology of tryptophan hydroxylase, the antibodies should cross-react with other mammalian species. Due to antigen modifications in E. coli, this antibody does not recognize TPH in rabbit tissues.
Polyclonal Sheep IgG
100 μL in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 μg/mL BSA and 50% glycerol.
Detection of Tryptophan Hydroxylase by Western Blot.
Western blot of human dorsal Raphe nucleus. As shown in the autoradiograph, the antibody is specific for the ~55 kDa TPH protein.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
For long-term storage, ≤ ‑20° C is recommended. Product is stable at ≤ ‑20° C for at least 1 year.
Background: Tryptophan Hydroxylase
Tryptophan hydroxylase (TPH) is a 51 kDa, 444 amino acid (aa) enzyme that belongs to the pterin-dependent, aromatic amino-acid hydroxylase family. It is found in peripheral tissues and pineal tissues, and catalyzes the rate-limiting step in serotonin generation from L‑tryptophan. The molecule contains two domains, an N-terminal regulatory domain (aa 1-110) and a C-terminal catalytic domain (aa 111-444) that contains an iron atom. TPH is constitutively active but unstable. Phosphorylation by PKA at S58 stabilizes the enzyme and increases its activity by 25%. This action is complemented by 14-3-3 proteins that physically interact with phosphorylated TPH to increase both its stability and activity by another 15% to 45% of baseline. Binding of the acidic carboxyterminus of 14-3-3 with the regulatory region of TPH also blocks phosphatase activity directed towards TPH, prolonging its activity. One splice variant of TPH is known. It occurs at the extreme C‑terminus where there is a 29 aa substitution for the last seven amino acids. This may generate a more stable enzyme.
Boularand, S. et al. (1990) Nucleic Acids Res. 18:4257.
Wang, G-A. et al. (1998) J. Neurochem. 71:1769.
Kumer, S.C. et al. (1997) J. Neurochem. 69:1738.
Wang, L. et al. (2002) Biochemistry 41:12569.
Banik, U. et al. (1997) J. Biol. Chem. 272:26219.
Haycock, J.W. et al. (2002) J. Neurosci. Methods. 114:205.
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