Technical Data
5-Hydroxytryptophan, Conjugated
Application(s): Suitable for use in Immunohistochemistry, ELISA. . Other applications not tested.

Recommended Dilution
Immunohistochemistry: 1:2000-1:5000. Tested using the free floating PAP technique on rat raphe nuclei. The anti-conjugated 5-hydroxytryptophan antibodies stains well.

Additional crossreactivity data:
Using a conjugate 5 hydroxytryptophan-Glutaraldehyde-Protein, antibody specificity was performed with an ELISA test by competition experiments with the following compounds:

Compounds Crossreactivity ratio(a)
5-Hydroxytryptophan-G-BSA 1
5-Methoxytryptophan-G-BSA 1:2000
Tryptophan-G-BSA 1:50,000
5-Hydroxytryptamine-G-BSA 1:50,000
Tryptamine-G-BSA 1:>50,000
5-Hydroxytryptophan 1:>50,000

(a): 5-Hydroxytryptophan-G-BSA concentration/unconjugated or conjugated indolealkylamine concentration at half displacement. G = Glutaraldehyde, BSA

PAP procedure:
Second antibody : Sections are incubated with 1/100 dilution of goat anti-rabbit in solution C for 3 hours at 20C or 1 hour at 37C. Then, they are washed 3 times (10 min) with solution C.

PAP : Sections are incubated with 1/1000 dilution of rabbit peroxidase anti-peroxidase complex in solution C for 1 hour at 37C. Then, they are washed 3 times (10 min) with solution C.

Antibody-antigen complexes are revealed using diaminobenzidine (25mg/100ml) (or other chromogen) dissolved in Tris 0.05M and filtrated ; 0.05% of H2O2 is added. The sections are incubated for 10 min at 20C. Reaction is stopped by transfering sections in 5ml of Tris 0.05M.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20C. Aliquots are stable for at least 12 months at -20C. 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.
100ug-20CBlue IceRabbit
5 Hydroxytryptophan Glutaraldehyde Carriers.
Antiserum previously preabsobed on protein carriers, and purified by ammonium sulfate precipitation and gel filtration.
Supplied as a liquid in PBS, pH 7.2. No preservative added.
Recognizes protein-conjugated 5-Hydroxytryptophan. See additional crossreactivity data.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
Geffard M., Touret M. and Kitahama K. First characterization of 5-hydroxytryptophan in rat brain by using specific antibodies, Brain Res., (1987), 426, 191-196.
Touret M., Kitahama K., Geffard M. and JOUVET M. 5-hydroxytryptophan (5-HTP)-immunoreactive neurons in the rat brain tissue. Neurosc. Letters, (1987) 80, 263-267.
Touret M., Sarda N., Gharib A., Geffard M. and Jouvet M. The role of 5-hydroxytryptophan (5-HTP) in the regulation of the sleep/wake cycle in parachlorophenylalanine (p-CPA) pretreated rat : a multiple approach study. Exp. Brain Res., (1991) 86, 117-124.