| Catalog # | H9119 |
| 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 20°C or 1 hour at 37°C. 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 37°C. 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 20°C. Reaction is stopped by transfering sections in 5ml of Tris 0.05M. |
| Storage and Stability | May be stored at 4°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°C. Aliquots are stable for at least 12 months at -20°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. |
| CAS Number | n/a |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in PBS, pH 7.2. No preservative added. |
| Purity | Antiserum previously preabsobed on protein carriers, and purified by ammonium sulfate precipitation and gel filtration. |
| Immunogen | 5 Hydroxytryptophan Glutaraldehyde Carriers. |
| Specificity | Recognizes protein-conjugated 5-Hydroxytryptophan. See additional crossreactivity data. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
