M1 mouse anti-FLAG monoclonal antibody, normal goat serum, poly-D-lysine, triton X-100, norbinaltorphimine (nor-BNI), glibenclamide, iberiotoxin, and sulpiride were obtained from Sigma-Aldrich (St. Louis, MO). Salvinorin A (SA, purity > 98%) was obtained from Apple Pharms (Asheville NC, USA). Alexa Fluor 488-conjugated goat anti-mouse immunoglobulin (IgG), Alexa Fluor 488-conjugated goat anti-rabbit IgG with 10 nm colloidal gold conjugate, Antifade Mounting Medium with DAPI, Minimum Essential Medium (MEM) powder, glucose-free Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS) were obtained from Invitrogen (Carlsbad, CA). Anti-RFP antibody and Dyngo-4a were obtained from Abcam (Cambridge, MA). Geneticin was obtained from Cellgro Mediatechc (Herndon, VA). Lab-Tek II Slide Chamber was obtained from Thermo Scientific (Waltham, MA). Hypoxia system with gas mixture (95% N2, 5% CO2) was obtained from Airgas USA, LLC (Cherry Hill, NJ). All other chemicals used were of reagent grade and obtained from Sigma-Aldrich.
Nor binaltorphimine nor bni
Manufactured by Merck Group
Sourced in United States
Nor-binaltorphimine (nor-BNI) is a selective kappa opioid receptor antagonist used in laboratory research. It is a chemical compound utilized for its pharmacological properties in the study of opioid receptor systems and their functions.
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Lab products found in correlation
Naltrindole, by Merck Group (2 mentions)
Cocaine hydrochloride, by Merck Group (2 mentions)
Sulpiride, by Merck Group (1 mentions)
Iberiotoxin, by Merck Group (1 mentions)
Glibenclamide, by Merck Group (1 mentions)
Normal goat serum (ngs), by Merck Group (1 mentions)
Anti rfp antibody, by Abcam (1 mentions)
Alexa fluor 488 conjugated goat anti mouse immunoglobulin igg, by Thermo Fisher Scientific (1 mentions)
Anti fade mounting medium with dapi, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Poly d lysine, by Merck Group (1 mentions)
Dyngo 4a, by Abcam (1 mentions)
Ana 12, by Merck Group (1 mentions)
U50 488, by Merck Group (1 mentions)
Esi q tof maxis 4g, by Bruker (1 mentions)
β fna, by Merck Group (1 mentions)
Sb203580, by Beyotime (1 mentions)
Fmoc protected amino acids, by GL Biochem (1 mentions)
Carrageenan, by Merck Group (1 mentions)
Tnf α, by Merck Group (1 mentions)
7 protocols using nor binaltorphimine nor bni
1
Immunofluorescence Imaging of G-Protein Signaling
2
Antagonizing BDNF and Kappa Opioid Receptors
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ANA-12, a BDNF receptor (tropomyosin-related kinase B (TrkB)) antagonist, and nor-binaltorphimine (nor-BNI), a selective κ-opioid receptor (KOR) antagonist, were purchased from Sigma Chemical (St. Louis, MO). ANA-12 and nor-BNI were dissolved in 0.9% saline. ANA-12 (100 nmol) or nor-BNI (5 nmol) administered to animals by intrathecal route once (5 µl) and testing done in 30–60 min. The selected drug doses and timing of administration were in accordance with prior publications.28 (link)–30 (link, link)
3
Isolation and Synthesis of Salvinorin A Analogs
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Salvinorin A was isolated and purified from Salvia divinorum leaves and assessed for purity (>98%) using high-performance liquid chromatography (HPLC). The SalA analogs, 16-Ethynyl SalA and 16-Bromo SalA, were synthesized as previously described (Riley et al., 2014 (link)) and were tested for purity (>95%) with HPLC. Nor-binaltorphimine (nor-BNI) and U50,488 were purchased from Sigma-Aldrich (St. Louis, MO, United States).
4
Solid-Phase Synthesis of EM-1 and EM-2 Peptides
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EM-1 and EM-2 were prepared by manual solid-phase synthesis using standard N-fluorenylmethoxycarbonyl (Fmoc) chemistry as reported in our previous study [32 (link)]. Fmoc-protected amino acids (GL Biochem Ltd., China) were coupled with Rink amide 4-methybenzhydrylamine (MBHA) resin (Tianjin Nankai Hecheng Science & Technology Co., Ltd., China). The crude peptides were purified by preparative reversed-phase HPLC (RP-HPLC) and determined by electrospray ionization mass spectrometer (ESI-Q-TOF maXis-4G, Bruker Daltonics).
Naloxone, β-FNA, nor-binaltorphimine (nor-BNI), and naltrindole (NTI) were obtained from Sigma-Aldrich. The selective p38 MAPK inhibitor SB203580 was purchased from Beyotime Institute of Biotechnology and dissolved in 1% DMSO in saline. All other drugs were dissolved in sterilized saline and stored at − 20 °C.
Naloxone, β-FNA, nor-binaltorphimine (nor-BNI), and naltrindole (NTI) were obtained from Sigma-Aldrich. The selective p38 MAPK inhibitor SB203580 was purchased from Beyotime Institute of Biotechnology and dissolved in 1% DMSO in saline. All other drugs were dissolved in sterilized saline and stored at − 20 °C.
5
Opioid and Cocaine Receptor Binding
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Norbinaltorphimine (nor-BNI) and cocaine hydrochloride were obtained from Sigma-Aldrich (St. Louis, MO, USA) and diluted to concentration in sterile saline (0.9%). Compound 1 was synthesized as described above and diluted to concentration in a vehicle (Solutol HS 30% in sterile 0.9% saline).
6
Modulation of Inflammatory Responses
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-Carrageenan, TNF-a, PGE 2 , dopamine (DA), ethylenediamine tetra acetic acid (EDTA), lip polysaccharide (LPS), Griess reagent, Tu ¨rk solution, and PC (99.7% purity) were purchased from Sigma (St. Louis, MO). Enzyme-linked immunosorbent assay (ELISA) for mouse quantitative determination of TNF-a was obtained from BD-Bioscience Pharmingen (San Diego, CA). Sodium nitrite (NaNO 2 ) was obtained from Merck, Darmstadt, Germany. Indomethacin and dipyrone were obtained from Unia ˜o Quı ´mica (Sa ˜o Paulo, Brazil). Naloxone (non-selective antagonist of opioid receptors), naltrindole (d-opioid receptor antagonist), and norbinaltorphimine (Nor-BNI; k-opioid receptor antagonist) were purchased from Sigma Chemical Company (St. Louis, MO). D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr amide (CTOP; m-opioid receptor antagonist) was purchased from Tocris Bioscience (Bristol, UK). Morphine was purchased from Crista ´lia (Itapira, Sa ˜o Paulo, Brazil). Drugs were dissolved in physiological saline and administered through intraperitoneal (i.p.) or subcutaneous (s.c.) routes.
7
Norbinaltorphimine and Cocaine Administration
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Norbinaltorphimine (nor-BNI) and cocaine hydrochloride were obtained from Sigma-Aldrich (St. Louis, MO, USA) and diluted to concentration in sterile saline (0.9%). 1 was synthesized as described above and diluted to concentration in the vehicle (solutol HS 30% in sterile 0.9% saline).
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