Jwh015

Manufactured by Merck Group

Sourced in United States

JWH015 is a laboratory research chemical compound primarily used in scientific research. It functions as a selective agonist for the CB2 cannabinoid receptor. The core function of JWH015 is to facilitate the study of the endocannabinoid system and its interactions.

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Lab products found in correlation

Sr144528, by Cayman Chemical (2 mentions) Am630, by Merck Group (2 mentions) Anandamide, by Cayman Chemical (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Rimonabant, by Cayman Chemical (1 mentions) Anti cb1, by Cayman Chemical (1 mentions) Protease phosphatase inhibitor cocktail, by Merck Group (1 mentions) Trypsin edta solution, by Merck Group (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Rpmi 1640 medium, by Merck Group (1 mentions) Sulfanilamide, by Merck Group (1 mentions) Win55 212 2, by Bio-Techne (1 mentions) Am630, by Bio-Techne (1 mentions) 35s gtpγs, by American Radiolabeled Chemicals (1 mentions) Am281, by Bio-Techne (1 mentions) Lipopolysaccharide from e coli 0111 b4, by Merck Group (1 mentions) Cp55940, by Merck Group (1 mentions) Ridaifen b, by Merck Group (1 mentions)

6 protocols using jwh015

Cannabinoid Receptor Signaling Modulation

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RPMI 1640 medium, trypsin-EDTA solution, MTT, protease/phosphatase inhibitor cocktail, and JWH015, were purchased from Sigma (Milan, IT). The fetal calf serum was from Gibco (Euroclone, Milan, IT). Primary antibodies anti-CB1, anti-CB2 and anti-FAAH, anandamide, rimonabant, SR144528, oleoyl ethyl amide and Annexin V/PI detection kit were from Cayman Chemical (Space Import, Milan, IT). CB2 gene silencing was carried out by using TriFECTa™ Dicer-Substrate RNAi kit (ID HSC.RNAi.N001841.12) according to manufacturer’s (Integrated DNA Technologies) directions. The primary antibody anti-GAPDH and HRP conjugated secondary antibodies were from Santa Cruz Biotechnology. Primary antibodies anti-cleaved Caspase-3, Ezrin EZR, pERM were from Cell Signaling Technology (Danvers, MA, USA), while anti SRC/pSRC were from AbCam. [1-³H]sphingosine and [³H]lipids used as chromatographic standards were kindly provided by Prof. Sonnino of the University of Milan.

Bettiga A., Aureli M., Colciago G., Murdica V., Moschini M., Lucianò R., Canals D., Hannun Y., Hedlund P., Lavorgna G., Colombo R., Bassi R., Samarani M., Montorsi F., Salonia A, & Benigni F. (2017). Bladder cancer cell growth and motility implicate cannabinoid 2 receptor-mediated modifications of sphingolipids metabolism. Scientific Reports, 7, 42157.

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Intrathecal CB2R Agonist and Antagonist Study

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The CB2R agonist JWH015 (2 µg; sigma, USA) and antagonist AM630 (1 µg; sigma, USA) were dissolved in 5 µL of 20% dimethyl sulfoxide (DMSO) (in normal saline). On day 14 after sarcoma cells implantation, mice in the tumor+JWH015 group received a single intrathecal injection of 2 µg of JWH015. Mice in the tumor+AM630+JWH015 group received a single intrathecal injection of 1 µg of AM630 30 min before the injection of JWH015. Mice in the tumor+vehicle group received the same volume of 20% DMSO under identical conditions as the control. Doses of these compounds were based on our previous study.18 (link)

Wang C., Xu K., Wang Y., Mao Y., Huang Y., Liang Y., Liu Y., Hao J., Gu X., Ma Z, & Sun Y. (2020). Spinal cannabinoid receptor 2 activation reduces hypersensitivity associated with bone cancer pain and improves the integrity of the blood–spinal cord barrier. Regional Anesthesia and Pain Medicine, 45(10), 783-791.

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Osteoclastogenesis Assay Protocol

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Ridaifen-B, CP-55,940, and JWH-015 were purchased from Sigma Aldrich (St. Louis, MO). WIN-55,212-2, AM-630, and AM-281 were obtained from Tocris Bioscience (Minneapolis, MN). SR-144528 was procured from Cayman Chemicals (Ann Arbor, MI). The radioligand [³H]CP-55,940 (131.4 Ci/mmol) was purchased from Perkin Elmer (Waltham, MA) and [³⁵S]GTPγS (1250 Ci/mmol) from American Radiolabeled Chemicals (St. Louis, MO). All compounds were diluted to 10⁻²M in 100% DMSO and stored at −20 °C. N-1 napthylethlene, sulfanilamide, and lipopolysaccharide from E. coli 0111:B4 were purchased from Sigma Aldrich (St. Louis, MO). ELISA kits to measure levels of mouse TNFα and mouse IL-6 were obtained from R&D systems (Minneapolis, MN). Mouse IL-1α release was quantified by ELISA kits procured from Ray Biotech (Norcross, GA). The WST-1 cell proliferation reagent from CellPro Roche was obtained from Sigma Adrich (St. Louis, MO). Reagents for TRAP staining were purchased from Sigma Aldrich (St. Louis, MO). All other supplies were purchased from Fisher Scientific.

Franks L.N., Ford B.M., Fujiwara T., Zhao H, & Prather P.L. (2018). The tamoxifen derivative ridaifen-B is a high affinity selective CB2 receptor inverse agonist exhibiting anti-inflammatory and anti-osteoclastogenic effects. Toxicology and applied pharmacology, 353, 31-42.

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Cannabinoid and Opioid Receptor Agonist Protocol

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JWH015, a cannabinoid 2 receptor (CB2) agonist (CB₂ K_i = 13.8 nM, 30 to 80 fold selectivity versus CB₁) was obtained from Cayman Chemical (Ann Arbor, MI). JWH015 was dissolved in a vehicle solution of 10% dimethyl sulfoxide, 10% Tween-80, and 80% saline (Sigma, St. Louis, MO). The mu-opioid agonist (MOR) morphine sulfate was purchased from the NIDA Drug Supply program (Rockville, MD) and was dissolved in saline. Formalin solution was obtained by diluting 1.5% of formaldehyde in saline. Ketamine:xylazine (80 mg/kg: 12 mg/kg; Phoenix Pharmaceutical, St. Joseph, MO) was used to anesthetize animals in order to insert microdialysis probes into the nucleus accumbens. The antibiotic gentamicin (Phoenix Pharmaceutical, St. Joseph, MO) was provided as a single subcutaneous dose (8mg/kg). Drugs were weighed out and dissolved in vehicle daily, prior to use. Cocaine hydrochloride was purchased from the NIDA Drug Supply program (Rockville, MD) and used as a positive control in microdialysis and HPLC studies. Finally, 5% and 2.5% isofluorane (Sigma, St. Louis, MO) mixed in 2.5 L/min of oxygen was delivered through a nose cone and used to induce and maintain anesthesia, respectively, for paw incision and SNI surgeries.

Grenald S.A., Young M.A., Wang Y., Ossipov M.H., Ibrahim M.M., Largent-Milnes T.M, & Vanderah T.W. (2016). Synergistic attenuation of chronic pain using mu opioid and cannabinoid receptor 2 agonists. Neuropharmacology, 116, 59-70.

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Glioma and Microglial Cell Culture

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IMG, C6 glioma, and BV-2 cells were cultured in Dulbecco’s modified Eagle medium (DMEM) with high glucose (4.5 g/L), 10 % fetal bovine serum (FBS) and penicillin/streptomycin (100 U/mL). SH-SY5Y cells were maintained in DMEM/F12 (1:1 ratio) media with 10 % FBS and penicillin/streptomycin (100 U/mL). IFNγ, IL-1β, TNF-α, IL-4, IL-6, and IL-13 were purchased from Peprotech (Rocky Hill, NJ). LPS, Ac-YVAD-CMK, delta-9-tetrahydrocannabinol, and JWH-015 were purchased from Sigma Aldrich. Adenosine triphosphate (ATP) was purchased from Amersham Biosciences.

McCarthy R.C., Lu D.Y., Alkhateeb A., Gardeck A.M., Lee C.H, & Wessling-Resnick M. (2016). Characterization of a novel adult murine immortalized microglial cell line and its activation by amyloid-beta. Journal of Neuroinflammation, 13, 21.

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CB2 Receptor Modulation in Neuroimmune Interactions

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For in vivo experiments, drugs were prepared and administered as previously described (29 (link),32 (link),33 (link)). The selective CB2 agonist JWH015 (Sigma-Aldrich; Merck KGaA) was dissolved in 5% DMSO corresponding to a dose of 1 µg/5 µl (50 µg/kg) or 2 µg/5 µl (100 µg/kg). The CB2-selective antagonist AM630 (Sigma-Aldrich; Merck KGaA) was dissolved in 5% DMSO corresponding to a dose of 2 µg/5 µl (100 µg/kg). Bafilomycin A1 (Baf-A1), an inhibitor of autophagosome and lysosome fusion, was dissolved in 5% DMSO corresponding to a dose of 10 nM. To avoid systemic effects on tumor cells, intrathecal administration was selected (34 (link)). Drugs were intrathecally administered at a volume of 5 µl at day 14 after the inoculation with tumor cells. To antagonize the activation of the CB2 receptor, AM630 was injected 30 min before JWH015.
For in vitro experiments, JWH015 and AM630 were dissolved in culture medium corresponding to a dose of 1 µM. Lipopolysaccharide (LPS) was dissolved in culture medium corresponding to a dose of 100 nM.

Mao Y., Huang Y., Zhang Y., Wang C., Wu H., Tian X., Liu Y., Hou B., Liang Y., Rong H., Gu X, & Ma Z. (2019). Cannabinoid receptor 2-selective agonist JWH015 attenuates bone cancer pain through the amelioration of impaired autophagy flux induced by inflammatory mediators in the spinal cord. Molecular Medicine Reports, 20(6), 5100-5110.

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