5-Hydroxytryptophan, Conjugated

Application(s): Suitable for use in Immunohistochemistry, ELISA. . Other applications not tested.\n\nRecommended Dilution\nImmunohistochemistry: 1:2000-1:5000. Tested using the free floating PAP technique on rat raphe nuclei. The anti-conjugated 5-hydroxytryptophan antibodies stains well.\n\nAdditional crossreactivity data:\nUsing a conjugate 5 hydroxytryptophan-Glutaraldehyde-Protein, antibody specificity was performed with an ELISA test by competition experiments with the following compounds: \n\nCompounds Crossreactivity ratio(a)\n5-Hydroxytryptophan-G-BSA 1\n5-Methoxytryptophan-G-BSA 1:2000\nTryptophan-G-BSA 1:50,000\n5-Hydroxytryptamine-G-BSA 1:50,000\nTryptamine-G-BSA 1:>50,000\n5-Hydroxytryptophan 1:>50,000\n\n(a): 5-Hydroxytryptophan-G-BSA concentration/unconjugated or conjugated indolealkylamine concentration at half displacement. G = Glutaraldehyde, BSA\n\nPAP procedure:\nSecond antibody : Sections are incubated with 1/100 dilution of goat anti-rabbit in solution C for 3 hours at 20 degrees C or 1 hour at 37 degrees C. Then, they are washed 3 times (10 min) with solution C.\n\nPAP : Sections are incubated with 1/1000 dilution of rabbit peroxidase anti-peroxidase complex in solution C for 1 hour at 37 degrees C. Then, they are washed 3 times (10 min) with solution C.\n\nAntibody-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 20 degrees C. Reaction is stopped by transfering sections in 5ml of Tris 0.05M. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. 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.

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SPECIFICATIONS

Catalog Number

H9119

Size

100ug

Applications

ELISA, IHC

Hosts

Rabbit

Reactivities

Rat

Form

Supplied as a liquid in PBS, pH 7.2. No preservative added.

P Type

Pab

Purity

Antiserum previously preabsobed on protein carriers, and purified by ammonium sulfate precipitation and gel filtration.

Isotype

IgG

References

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.\nTouret 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.\nTouret 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.

Additional Info

Recognizes protein-conjugated 5-Hydroxytryptophan. See additional crossreactivity data.