Detection of Phospho-Tryptophan Hydroxylase 1/TPH‑1 (S260) by Western Blot. Western blot of rat brain stem lysate showing specific immunolabeling of the approximately 51 kDa TPH phosphorylated at S260. The labeling is specifically blocked by the S260 phosphopeptide (Peptide block) used as antigen. The corresponding non-phosphopeptide (Negative control) did not block the immunolabeling.
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 1/TPH-1
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 gland, 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 CaMPKII occurs at S58 and S260. The effects on activity are unclear. PKA phosphorylation 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. Presumably, CaMPKII activity at S58 would have the same effect. An equivalent situation has not been reported for S260. 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.
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