The following were purchased from Cayman (Ann Arbor, MI): 2-AG; the endocannabinoid receptor inhibitors, SR141716A (Rimonabant, CB1 inverse agonist), AM251 (CB1 antagonist) and SR144528 (CB2 inverse agonist); the endocannabinoid receptor activators, CP47497 (CB1 agonist), AM1241 (CB2 agonist), Win- 55–212-2 (CB1 and CB2 agonist) and CP55940 (CB1 and CB2 agonist); the selective and nonselective inhibitors for cyclooxygenase-1 and -2 (COX-1, COX-2), SC-560 (selective for COX-1), CAY10404 (selective for COX- 2), Ibuprofen (nonselective for COX-1 and COX-2); and the selective inhibitors for hydrolases of 2-AG, JZL 184 (selective for enzyme monoacylglycerol lipase, MAGL. Anti-bromodeoxyurine (BrdU) antibody was purchased from Roche Applied Science (Indianapolis, IN). The CellTiter 96 Non-Radioactive Cell proliferation Assay Kit™ was purchased from Promega (Madison, WI). Charcoal/dextran-treated fetal bovine serum (CFBS) was purchased from Hyclone (Logan, UT). DMEM, RPMI-1640, antibiotics (penicillin and streptomycin), and fetal bovine serum (FBS) were purchased from Atlanta Biologicals (Lawrenceville, GA).
Cay10404
Manufactured by Cayman Chemical
Sourced in United States
CAY10404 is a laboratory reagent provided by Cayman Chemical. It is a crystalline solid powder. The core function of this product is to serve as a chemical standard or reference material for research and analytical purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ns 398, by Cayman Chemical (4 mentions)
Celltiter 96 non radioactive cell proliferation assay kit, by Promega (1 mentions)
Jzl184, by Cayman Chemical (1 mentions)
Dulbecco modified eagle medium (dmem), by Atlanta Biologicals (1 mentions)
Am251, by Cayman Chemical (1 mentions)
Sr141716a, by Cayman Chemical (1 mentions)
Am1241, by Cayman Chemical (1 mentions)
Rpmi 1640, by Atlanta Biologicals (1 mentions)
Win55 212 2, by Cayman Chemical (1 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)
Sb 505124, by Merck Group (1 mentions)
Il 1ra, by Thermo Fisher Scientific (1 mentions)
Rabbit anti phospho smad3 antibody, by Rockland Immunochemicals (1 mentions)
Recombinant human il 1α, by Thermo Fisher Scientific (1 mentions)
Recombinant human tgf β1, by R&D Systems (1 mentions)
Isqavhaahaeineagr, by Merck Group (1 mentions)
Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Tumor necrosis factor α tnf α, by Merck Group (1 mentions)
9 protocols using cay10404
1
Endocannabinoid Receptor Modulation in Cell Proliferation
2
Influenza Virus Infection Kinetics
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A549 cells were pre-incubated with 5 μg/ml PGE2 (Sigma-Aldrich), diluted in ethanol, or 5 μM COX-2 inhibitor CAY10404 (Cayman Chemicals), diluted in DMSO or the respective solvent as control for 1 h. Subsequently, cells were infected with IAV in addition to the treatment with PGE2, CAY10404 or the respective solvent for 24 h. Viral titres were determined by Standard Plaque Titration Assay.
In time-of-addition kinetics A549 were either pre-incubated with 5 μM CAY10404, or 5 μg/ml PGE2 or the respective solvent for 1 h prior infection or left untreated. Subsequently, cells were infected with FPV (MOI 0.01). Substances were added at the indicated times of infection and viral titres were determined by Standard Plaque Titration Assay 10 h p.i.
In time-of-addition kinetics A549 were either pre-incubated with 5 μM CAY10404, or 5 μg/ml PGE2 or the respective solvent for 1 h prior infection or left untreated. Subsequently, cells were infected with FPV (MOI 0.01). Substances were added at the indicated times of infection and viral titres were determined by Standard Plaque Titration Assay 10 h p.i.
3
Investigating Cell Signaling Pathways
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Recombinant human TGF-β1 (R&D Systems, Minneapolis, MN, USA), recombinant human IL-1α (PeproTech, London, UK), and prostaglandin E2 (PGE2; Cayman Chemical Co., Ann Arbor, MI, USA) were used in the present experiment. The selective inhibitors used were an IL-1 receptor antagonist (IL-1Ra; PeproTech), TGF-β receptor inhibitor (SB-505124; Sigma-Aldrich), and selective cyclooxygenase-2 (COX-2) inhibitor (CAY10404; Cayman Chemical Co.). The antibodies used were mouse monoclonal anti-TGF-β1,2,3 antibody (clone 1D11; R&D Systems), mouse monoclonal anti-human RANKL antibody (clone 70525.11; Sigma-Aldrich), goat anti-human COX-2 antibody (Cayman Chemical Co.), and rabbit anti-phospho-Smad3 antibody (Rockland, Gilbertsville, PA, USA).
4
Kupffer Cell-Mediated CD4+ T Cell Modulation
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FACS-sorted Kupffer cells from control or HCC-bearing mice were cultured in 96-well plates overnight. Splenocytes and lymph node cells from OT-II mice were labeled with CFSE (0.5 µM). CD4+ T cells were purified using a CD4+ T Cell Isolation Kit (Miltenyi Biotec). After 3-h-incubation with ovalbumin323-339 peptide (ISQAVHAAHAEINEAGR, 40 ng/mL, Sigma-Aldrich) with or without LPS (400 ng/mL, Sigma-Aldrich), CFSE-labeled CD4+ cells were added to each well at different Kupffer cells/CD4+ cells ratio. For selected experiments anti-PD-L1 (10F.9G2), anti-IL-10 (JES5-16E3) (Biolegend), and CAY10404 (Cayman Chemical) were added.
5
Isolation and Characterization of NPC Cancer Stem-like Cells
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It has been reported that cancer stem-like SP cells were isolated from CNE1 and CNE2 10 (link), so CNE1 and CNE2 cells were used in the present study. (Of human NPC cell lines, CNE1 is well-differentiated, and CNE2 is poorly-differentiated). CNE1, CNE2, and 293T cells were obtained from the Cancer Center of Sun Yat-sen University (Guangzhou, China). The cells were maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) (Hyclone, UT, USA), 100 U/mL penicillin G, and 100 μg/mL streptomycin in a humidified incubator at 37°C with 5% CO2. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), Hoechst 33342, verapamil, dimethyl sulfoxide (DMSO), parthenolide (PN), pyrrolidine dithiocarbamate (PDTC), tumor necrosis factor-α (TNF-α), and 5-fluorouracil (5-FU) were from Sigma-Aldrich (MI, USA). The COX-2 inhibitors NS-398 and CAY10404 were purchased from Cayman Chemical (MI, USA). TRIzol was purchased from Invitrogen (CA, USA).
6
Quantification of Lipid Mediators
7
Lipid Metabolism and Signaling Inhibitor Protocols
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FA-free BSA, palmitic acid (PA), oleic acid (OA), AA, EPA, rosiglitazone, bortezomib, MG132, bafilomycin A1, and inhibitors to CYP2C and 2J (danazol and gemfibrozil) were purchased from Sigma. [9,10-3H(N)]PA (30–60 Ci; 1.11–2.22 TBq/mmol), [9,10-3H(N)]OA (15–60 Ci; 0.555–2.22 TBq/mmol), and [5,6,8,9,11,12,14,15-3H(N)]AA were obtained from Perkin-Elmer (Waltham, MA). Selective inhibitors of COX1 (SC-560), COX2 (CAY10404), and nonselective COX inhibitors (indomethacin and aspirin), 12- and 5-LOX inhibitor (3,4-dihydroxyphenyl ethanol, 5-LOX inhibitor (Zileuton), 15-LPX inhibitor 1, CYP4A, CYP4F inhibitor (HET0016), CYP2C9, 2C19, 3A inhibitor (fluconazole), and PGE2 were obtained from Cayman Chemicals (Ann Arbor, MI). Specific kinase inhibitors were purchased from Calbiochem (Temecula, CA). All other reagents used were of analytical grade.
8
Modulating Prostaglandin E2 Signaling in Hematopoietic Cells
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Cells were cultured with pharmacological compounds or antibodies to block PGE2 production and/or signaling by incubation with 10 µM indomethacin, 10 µM NS-398, 1 µM CAY10404 (Cayman Chemical), or 1:1,000 anti-PGE2 antibodies (Abcam) for the duration of culture with or without WSS. H89 (Cayman Chemical) was applied at 10 µM to inhibit PKA activity. Forskolin (Sigma-Aldrich) was administered at 10 µM as an adenylyl cyclase activator to stimulate cAMP production. The stabilized synthetic analogue dmPGE2 (Cayman Chemical) was applied to freshly dissociated AGM cells for 2 h at 37°C from 1 to 100 µM for colony formation assays or at 10 µM for transplantation into adult recipient mice. Intracellular calcium was sequestered by 10 µM BAPTA-AM (Tocris Bioscience) and elevated by 2 µM of calcium ionophore A23187 (Tocris Bioscience).
9
Kainic Acid and BzATP-Induced Inflammatory Response
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Tissue culture media and reagents along with the kainic acid and 3′-O-(Benzoyl) benzoyl ATP (BzATP) were purchased from Sigma Chemical Company (Saint Louis, MO). Recombinant mouse TNFα was purchased from R&D systems (Minneapolis, MN). Fetal bovine serum and horse serum were purchased from Hyclone (Logan, UT). All the COX-2 inhibitors (CAY 10452, NS398, and CAY 10404) and the EP2 agonist butaprost were purchased from Cayman Chemical Company (Ann Arbor, MI). The EP3 antagonist ONO-AE5-599 was provided by Ono Pharmaceuticals.
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