proDynorphin Catalog Number: Product Type: Immunogen Sequence: Data Sheet Affinity Purified SQENPNTYSEDLDV. Corresponding to residues 235-248 of the rat proDynorphin. Species Reactivity: Format: Rat, Mouse PBS liquid containing 0.05% sodium azide . Concentration 1 mg/ml Immunohistochemistry 1:250-1:1000* Dilutions listed as a recommendation. Optimal dilution should be determined by investigator. Maintain at +2-8 C for 3 months or at -20 C for longer periods. Stable for 1 year. Avoid repeated freeze-thaw cycles. B.A.S. Reyes, A.F. Carvalho, K. Vakharia. and E.J. Van Bockstaele. Amygdalar peptidergic circuits regulating noradrenergic locus coeruleus neurons: Linking limbic and arousal centers. doi:10.1016/j.expneurol.2011.04.001. Wulf Haubensak, Prabhat S. Kunwar, Haijiang Cai1, Stephane Ciocchi, Nicholas R. Wall, Ravikumar Ponnusamy, Jonathan Biag, Hong-Wei Dong, Karl Deisseroth, Edward M. Callaway, Michael S. Fanselow, Andreas Lüthi & David J. Anderson. Genetic dissection of an amygdala microcircuit that gates conditioned fear. Nature 468, 270-276 (11 November 2010) | doi:10.1038/nature09553. Melissa L. Perreault, Ahmed Hasbi, Mohammed Alijaniaram, Theresa Fan, George Varghese, Paul J. Fletcher, Philip Seeman, Brian F. O'Dowd, and Susan R. George. The Dopamine D1D2 Receptor Heteromer Localizes in Dynorphin/Enkephalin Neurons: INCREASED HIGH AFFINITY STATE FOLLOWING AMPHETAMINE AND IN SCHIZOPHRENIA. J. Biol. Chem., Nov 2010; 285: 36625 - 36634. Cristina Núñez, Fátima Martín, Anna Földes, M. Luisa Laorden, Krisztina J. Kovács, M. Victoria Milanés. Induction of FosB/ΔFosB in the brain stress system-related structures during morphine dependence and withdrawal. Journal of Neurochemistry. DOI: 10.1111/j.14714159.2010.06765.x N. Maolood and B. Meister. Nociceptin/Orphanin FQ Peptide in Hypothalamic Neurones Associated with the Control of Feeding Behaviour. Journal of Neuroendocrinology. Volume 22 Issue 2, Pages 75 - 82. Published Online: 15 Dec 2009. Application Notes Immunohistochemistry: Antiserum was used on perfusion fixed tissue. Perfusion: 1) calcium-free Tyrode’s solution, 2) paraformaldehyde-picric acid fixative, and 3) 10% sucrose in PBS as a cryo-protectant. Desired tissues were dissected and stored overnight in 10% sucrose in PBS. 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. V308/2011 Neuromics Antibodies • 5325 West 74 Street, Suite 8 • Edina, MN 55439 phone 866-350-1500 • fax 612-677-3976 • e-mail pshu

Slide-mounted tissue sections were processed for indirect immunofluorescence. Slides were incubated with blocking buffer for 1 hour at room temperature. Primary antiserum was diluted with blocking buffer to the appropriate working concentration. Blocking buffer was removed and slides were incubated for 18-24 hours at 4ºC with primary antiserum. Slides were rinsed 3 times and then incubated with secondary antibodies for 1 hour at room temperature. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy. Note: Sodium azide (NaN3) interferes with peroxidase...