Immunofluorescent analysis of TPH1 (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a TPH1 polyclonal antibody (Product # PA1-777) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse, Non-human primate, Rabbit, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the C-terminal region of TPH1.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:100-1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA1-777 detects TPH1 in human, mouse, rat, rabbit and non-human primate samples.
PA1-777 has been successfully used in ICC/IF, IP, and Western blot procedures. By Western blot PA1-777 detects a 51 kDa protein representing TPH1.
The PA1-777 immunogen is a synthetic peptide corresponding to the C-terminal region of TPH1.
Tryptophan hydroxylase (TPH) catalyzes the 5-hydroxylation of tryptophan, which is the first step in the biosynthesis of indoleamines (serotonin and melatonin) (Martinez et al., 2001). In mammals, serotonin biosynthesis occurs predominantly in neurons which originate in the Raphe nuclei of the brain, and melatonin synthesis takes place within the pineal gland. Although TPH catalyzes the same reaction within the Raphe nuclei and the pineal gland, TPH activity is rate-limiting for serotonin but not melatonin biosynthesis.
Serotonin functions mainly as a neurotransmitter, whereas melatonin is the principal hormone secreted by the pineal gland. The activity of TPH is enhanced by phosphorylation by cAMP-dependent protein kinase (PKA) and Ca2+/calmodulin kinase II (CaM K II) (Jiang et al., 2000; Johansen et al., 1996). CaM K II phosphorylates Ser260 which lies within the regulatory domain of TPH (Jiang et al., 2000).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Differential tissue distribution of tryptophan hydroxylase isoforms 1 and 2 as revealed with monospecific antibodies.
PA1-777 was used in western blot to investigate the localization of TPH isoforms in various tissues
|Sakowski SA,Geddes TJ,Thomas DM,Levi E,Hatfield JS,Kuhn DM||Brain research (1085:11)||2006|