5HT (High Titer) Catalog Number: Product Type: Immunogen Sequence: Applications: Data Sheet Host: Species Reactivity: Format: Goat Human, Mouse, Rat 100 ul. Lyophilized Immunohistochemistry: Recommended Dilution: 1/400 - 1/800 in PBS/0.3% Triton X-100 – Cy3 Technique; 1/5000 1/10,000 in PBS/0.3% Triton X-100 – Bn/Av-HRP Western Blot: Recommended Dilution: 1/500 or greater Dilutions listed as a recommendation. Optimal dilution should be determined by investigator. Storage: Dilute with phosphate buffer or Tris buffer at dilutions no higher than 1/10, aliquot and freeze at -15° C or lower. Antibody can be stored for up to six months if handled as described above. It is strongly recommended that the customer perform a primary antibody dilution series using our dilution recommendations as a guideline. Note that a change in the fixation or buffering system as used in our protocol may change the configuration Application Notes Immunohistochemistry Antigen: serotonin coupled to BSA with paraformaldehyde. Raised in goat Control Tissue: Rat hypothalamus, raphe nuclei and spinal cord. Perfusion Fixation: Fixative - 4% paraformaldehyde in 0.1 M Phosphate uffer, pH 7.4; 500 mL over approximately 20 minutes. Post Fixation: 4% paraformaldehyde in 0.1 M Phosphate buffer, pH 7.4; 500 mL over approximately 20 minutes. Post Fixation - 1.5 hour at 4° C. in 4% paraformaldehyde in 0.1 M phosphate buffer, pH 7.4. Note: Paraformaldehyde is a necessary component in fixation of serotonin. If needed, low levels of glutaraldehyde (0.10.3%) may be used in conjunction with paraformaldehyde. Sections: 10 μm cryostat or 50 μm vibratome Antibody dilution: 1/400-1/800 in PBS/0.3% Triton X-100 - Cy3 Technique; 1/5,000-1/10,000 in PBS/0.3% Triton X-100 - BnSA/HRP Technique Absorption control: 5-HT/BSA 25 μg/mL diluted serum. Incubation on Tissue: 16-24 hours at 4° C. Detection System: use Cy3 and Bn-SA/HRP at dilutions recommended by the manufacturers. Image: 5HT Staining of rat hypothalamus. FOR RESEARCH USE ONLY NEUROMICS’ REAGENTS ARE FOR IN VITRO AND CERTAIN NON-HUMAN IN VIVO EXPERIMENTAL USE ONLY AND NOT INTENDED FOR USE IN ANY HUMAN CLINICAL INVESTIGATION, DIAGNOSIS, PROGNOSIS, OR TREATMENT. THE ABOVE ANALYSES ARE MERELY TYPICAL GUIDES. THEY ARE NOT TO BE CONSTRUED AS BEING SPECIFICATIONS. ALL OF THE ABOVE INFORMATION IS, TO THE BEST OF OUR KNOWLEDGE, TRUE AND ACCURATE. HOWEVER, SINCE THE CONDITIONS OF USE ARE BEYOND OUR CONTROL, ALL RECOMMENDATIONS OR SUGGESTIONS ARE MADE WITHOUT GUARANTEE, EXPRESS OR IMPLIED, ON OUR PART. WE DISCLAIM ALL LIABILITY IN CONNECTION WITH THE USE OF THE INFORMATION CONTAINED HEREIN OR OTHERWISE, AND ALL SUCH RISKS ARE ASSUMED BY THE USER. WE FURTHER EXPRESSLY DISCLAIM ALL WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE. V2-08/2013 www.neuromics.com Neuromics Antibodies • 5325 West 74th Street, Suite 8 • Edina, MN 55439 phone 866-350-1500 • fax 612-677-3976 • e-mail pshus