Drugs and antibodies were administered as described in the main text. Indomethacin, SC-236 and SC-560 were purchased from Sigma (St. Louis, MO), solvated in 1% Tween 80 in PBS, and 100 μL of each drug suspension or buffer alone per 20 mg body weight was administered by oral gavage. Clodronate and PBS liposomes were manufactured as previously described [32 (link), 33 (link)]. Rat anti-C-C chemokine receptor type 2 (CCR2) (clone MC-21) was generated as previously described [34 (link), 35 (link)]. Rat anti-lymphocyte antigen 6G (Ly6G) (clone 1AG) and rat IgG2a and IgG2b isotype controls were purchased from Bio X Cell (West Lebanon, NH). All antibodies were diluted in tissue culture grade sterile Dulbecco’s PBS and 200 μL of each antibody solution were injected intraperitoneally as indicated.
Sc 236
Manufactured by Merck Group
Sourced in Sao Tome and Principe
SC-236 is a piece of laboratory equipment designed for conducting scientific research and analysis. It is a versatile instrument that can be used for a variety of applications. The core function of SC-236 is to perform accurate and precise measurements and data collection, without making any claims about its intended use or applications.
Automatically generated - may contain errors
Lab products found in correlation
Sc 560, by Merck Group (4 mentions)
Indomethacin, by Merck Group (3 mentions)
Rat anti lymphocyte antigen 6g ly6g clone 1ag, by BioXCell (2 mentions)
Rat igg2a and igg2b isotype controls, by BioXCell (2 mentions)
Goat pab to mouse igg, by Abcam (1 mentions)
Clofibrate, by Merck Group (1 mentions)
Goat pab to rabbit igg, by Abcam (1 mentions)
Palmitic acid, by Merck Group (1 mentions)
17 octadecynoic acid 17 odya, by Cayman Chemical (1 mentions)
Pgf2α, by Merck Group (1 mentions)
Pd98059, by Enzo Life Sciences (1 mentions)
Gf109203x, by Enzo Life Sciences (1 mentions)
Baicalein, by Cayman Chemical (1 mentions)
15 deoxy δ12 14 pgj2, by Merck Group (1 mentions)
U0126, by Enzo Life Sciences (1 mentions)
Bapta am, by Bio-Techne (1 mentions)
7 protocols using sc 236
1
Pharmacological Modulation of Inflammation
2
Pharmacological Modulation of Immune Responses
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Drugs and antibodies were administered as described in the main text. Indomethacin, SC-236 and SC-560 were purchased from Sigma (St. Louis, MO), solvated in 1% Tween 80 in PBS, and 100 μl of each drug suspension or buffer alone per 20 mg body weight was administered by oral gavage. Clodronate and PBS liposomes were manufactured as previously described (Seiler et al., 1997 (link), Zeisberger et al., 2006 (link)). Rat anti-C-C chemokine receptor type 2 (CCR2) (clone MC-21) was generated as previously described (Mack et al., 2001 (link), Bruhl et al., 2007 (link)). Rat anti-lymphocyte antigen 6G (Ly6G) (clone 1AG) and rat IgG2a and IgG2b isotype controls were purchased from Bio X Cell (West Lebanon, NH). All antibodies were diluted in tissue culture grade sterile Dulbecco's PBS and 200 μl of each antibody solution were injected intraperitoneally as indicated.
3
Lipid Metabolism Regulation Assay
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Chemicals used include SB 204990 (Thermo Fisher Scientific, Paisley, UK), TOFA, C75, clofibrate, palmitic acid, and SC-236 (Sigma-Aldrich, Dorset, UK), PGD2, PGI2, and PGE2 (Cambridge Bioscience, Cambridge, UK), and SC-51322, TG-4-155, and ER-819762 (Bio-Techne Ltd., Abingdon, UK), and all were reconstituted in dimethyl sulfoxide. TXA2 (Cambridge Bioscience, Cambridge, UK) was reconstitute in ethanol. PGF2α (Cambridge Bioscience, Cambridge, UK) and etomoxir and malonyl-CoA (Sigma-Aldrich, Dorset, UK) were reconstituted in E199 medium. Anti-β-actin mouse monoclonal antibody (MAb), anti-FASN goat polyclonal antibody (pAb), anti-COX2 goat pAb, and anti-ACC rabbit MAb (Abcam, Cambridge, UK) were used for Western blot analysis. Donkey pAb to goat IgG, goat pAb to mouse IgG, and goat pAb to rabbit IgG were also purchased from Abcam, Cambridge, UK.
4
Indirect Effects of MSU Crystals on Osteoblasts
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The collection of human bone samples was approved by the New Zealand Ministry of Health and Disability Ethics Committee (HDEC approval no. NTX/05/06/058). All study participants provided their written informed consent.
Cultures of primary osteoblasts (HOBs) were prepared from normal human trabecular bone obtained from patients undergoing knee or hip arthroplasty, using a method based on Robey and Termine [8 (link)]. For the study of the indirect effect of MSU crystals on HOBs, cells were seeded in 6-well plates at 5 × 104 cells/well in DMEM with 5% FBS and 10 µg/mL AA2P. The following day, media were changed to DMEM with 0.1% BSA and 10 µg/mL AA2P. Media were refreshed on the following day, and conditioned media from MSU crystal-stimulated THP-1 cells and controls were added, at 40% of the final volume. Cells were harvested and media collected prior to the addition of THP-1 conditioned media and following 2 h and 20 h of incubation. For the study of the role of COX-2 activation in the indirect effect of MSU crystals in HOBs, 1 μM of the COX-2-selective inhibitor (SC-236, Sigma-Aldrich) was added to HOBs for 1 h prior to the addition of conditioned medium from THP-1 cells. An equal volume of the SC-236 vehicle, DMSO, was added to the control wells.
Cultures of primary osteoblasts (HOBs) were prepared from normal human trabecular bone obtained from patients undergoing knee or hip arthroplasty, using a method based on Robey and Termine [8 (link)]. For the study of the indirect effect of MSU crystals on HOBs, cells were seeded in 6-well plates at 5 × 104 cells/well in DMEM with 5% FBS and 10 µg/mL AA2P. The following day, media were changed to DMEM with 0.1% BSA and 10 µg/mL AA2P. Media were refreshed on the following day, and conditioned media from MSU crystal-stimulated THP-1 cells and controls were added, at 40% of the final volume. Cells were harvested and media collected prior to the addition of THP-1 conditioned media and following 2 h and 20 h of incubation. For the study of the role of COX-2 activation in the indirect effect of MSU crystals in HOBs, 1 μM of the COX-2-selective inhibitor (SC-236, Sigma-Aldrich) was added to HOBs for 1 h prior to the addition of conditioned medium from THP-1 cells. An equal volume of the SC-236 vehicle, DMSO, was added to the control wells.
5
Selective COX Inhibition in Uroepithelial Cells
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Specific COX-1 inhibitor, SC-560 (1µM) and COX-2 inhibitor, SC-236 (5µM) (Sigma) were dissolved in DMSO and used in all the experiments. Equivalent volume of DMSO alone served as the vehicle control. Above used concentrations were not cytotoxic for human uroepithelial cells, 5637 and T-24 as confirmed by XTT cell viability assay [19 ] (Supplementary Fig. 1 ).
6
Investigating COX-2 Inhibition in Acute Cystitis
7
Protocols for Prostaglandin Pathway Modulation
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Carbaprostacyclin (cPGI2), AL 8810, baicalein, and 17-octadecynoic acid (17-ODYA) were purchased from Cayman Chemical. 15-Deoxy-Δ12,14 PGJ2, PGE2, PGF2α, SC-560, SC-236, and indomethacin were all purchased from Sigma-Aldrich. PD98059, U0126, and GF109203X were obtained from Enzo Life Sciences and BAPTA-AM from Tocris Bioscience.
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