Detection of Human Tryptophan Hydroxylase 1/TPH‑1 by Western Blot.
Western blot shows lysates of AGS human gastric adenocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Tryptophan Hydroxylase 1/TPH‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5276) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Tryptophan Hydroxylase 1 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Tryptophan Hydroxylase 1/TPH‑1 in Human Brain.
Tryptophan Hydroxylase 1/TPH‑1 was detected in immersion fixed paraffin-embedded sections of human brain (substantia nigra) using Goat Anti-Human Tryptophan Hydroxylase 1/TPH‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5276) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Tryptophan Hydroxylase 1/TPH‑1 in Rat Brain.
Tryptophan Hydroxylase 1/TPH‑1 was detected in perfusion fixed frozen sections of rat brain using Goat Anti-Human Tryptophan Hydroxylase 1/TPH‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5276) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to neurons. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Tryptophan Hydroxylase 1/TPH-1
Tryptophan Hydroxylase 1 (TPH-1) is a 50‑60 kDa intracellular enzyme that belongs to the biopterin-dependent, aromatic amino acid hydroxylase family. It is expressed in gut enterochromaffin cells and pineal gland. TPH-1 catalyzes the hydroxylation of L-Trp, which is the rate-limiting step in serotonin biosynthesis. Human TPH-1 is 444 amino acids (aa) in length. It contains an N-terminal regulatory domain (aa 1‑98), a catalytic region (aa 99‑424), and a C‑terminal Leu-zipper tetramerization domain (aa 425‑444). TPH-1 is phosphorylated on Ser58, and subsequently binds to 14-3-3 proteins, resulting in activation. It functions as a noncovalent Fe-binding 230 kDa homotetramer. There are three potential splice variants. One shows a 29 aa substitution for aa 438‑444, a second shows a 50 aa substitution for aa 157‑444, and a third shows a 68 aa substitution for aa 1‑21. Over aa 102‑403, Human TPH-1 shares 95% aa identical with mouse TPH-1.
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