Acetylcholine (ACh, #A6625), indomethacin (indo, #I8280), PEG-catalase (cat, #C4963), sodium nitroprusside (SNP, #71778), SP600125 (#S5567), tamoxifen (#T5648) and TCS401 (#SML2140) were purchased from Sigma Aldrich (St Louis, MO, US). l -NAME (N(γ)-nitro-l -arginine methyl ester, #06-651-00), phenylephrine (#18-605-133), SB203580 (#12-021-0), streptozotocin (#57-220-1), thapsigargin (Thapsi, #50-464-293), Tauroursodeoxycholic Acid (TUDCA, #NC1266953), and tunicamycin (Tunica, #35-161-0) were purchased from Fisher Scientific (Waltham, MA, US). Tempol (#ALX-430-081-G001) was purchased from Enzo life sciences (Farmingdale, NY, YS).
Indomethacin indo
Manufactured by Merck Group
Sourced in United States
Indomethacin (Indo) is a non-steroidal anti-inflammatory drug (NSAID) that can be used as a laboratory equipment for research purposes. It is a white to off-white crystalline substance that is practically insoluble in water. Indomethacin is commonly used in scientific research to study its pharmacological effects and mechanisms of action.
Automatically generated - may contain errors
Lab products found in correlation
Sodium nitroprusside snp, by Merck Group (6 mentions)
Streptozotocin (stz), by Thermo Fisher Scientific (2 mentions)
Tunicamycin, by Thermo Fisher Scientific (2 mentions)
Peg catalase, by Merck Group (2 mentions)
Tcs401, by Merck Group (2 mentions)
L name n γ nitro l arginine methyl ester, by Thermo Fisher Scientific (2 mentions)
Sb203580, by Thermo Fisher Scientific (2 mentions)
Thapsigargin thapsi, by Thermo Fisher Scientific (2 mentions)
Tauroursodeoxycholic acid, by Thermo Fisher Scientific (2 mentions)
Tempol, by Enzo Life Sciences (2 mentions)
Sp600125, by Merck Group (2 mentions)
Tamoxifen, by Merck Group (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Nω nitro arginine methyl ester l name, by Merck Group (2 mentions)
Lipopolysaccharides (lps), by Merck Group (2 mentions)
Nω nitro l arginine methyl ester l name, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Horseradish peroxidase hrp, by Solarbio (1 mentions)
Ussing chamber, by Physiologic Instruments (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
17 protocols using indomethacin indo
1
Pharmacological Modulation of Cellular Processes
2
Measuring Intestinal Barrier Function
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Colonic tissue was opened along the mesenteric border and mounted in Ussing chambers (Physiologic Instruments, San Diego, CA) with an 0.5 cm2 exposing area of the tissue surface to 5 mL of V O2/V CO2 95:5 Krebs-glucose (10 mM) at 37°C. After adapted for 20 minutes, tetrodotoxin (TTX, 0.1 μM, Shanghai Aladdin Biochemical Technology Co., Ltd., Shanghai, China) and indomethacin (Indo, 1μM, Sigma-Aldrich Corp., St. Louis, MO, USA) were added to the basal side, respectively, to inhibit enteric neurons and prostaglandin synthase and sections let they reached a steady-state level for at least 30 minutes before each experiment began.
The paracellular pathway and transcellular pathway were measured as the flux of 4-kDa FITC-dextran (FD-4; 0.5 mg/mL; Sigma-Aldrich Corp., St. Louis, MO, USA) and horseradish peroxidase (HRP; 0.5 mg/mL; Beijing Solarbio Science & Technology Co., Ltd., Beijing, China), respectively. FD-4 and HRP were added to the luminal side, and the samples of 400 μL were collected from the basolateral per 20 min for 2 h and replaced by an equal volume of the corresponding buffer. The concentration of FD-4 was measured using fluorescence at emission 528 nm and excitation 485 nm. 3.3′,5,5′-Tetramethylbenzidine (TMB) was used to detect HRP at absorbance 405 nm.18 (link)
The paracellular pathway and transcellular pathway were measured as the flux of 4-kDa FITC-dextran (FD-4; 0.5 mg/mL; Sigma-Aldrich Corp., St. Louis, MO, USA) and horseradish peroxidase (HRP; 0.5 mg/mL; Beijing Solarbio Science & Technology Co., Ltd., Beijing, China), respectively. FD-4 and HRP were added to the luminal side, and the samples of 400 μL were collected from the basolateral per 20 min for 2 h and replaced by an equal volume of the corresponding buffer. The concentration of FD-4 was measured using fluorescence at emission 528 nm and excitation 485 nm. 3.3′,5,5′-Tetramethylbenzidine (TMB) was used to detect HRP at absorbance 405 nm.18 (link)
3
Preparation and Use of Krebs-Ringer Buffered Solutions
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Krebs-Ringer bicarbonate-buffered physiological salt buffer (KRB-buffer) contained 118.5 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgSO4, 1.2 mM KH2PO4, 25.0 mM NaHCO3 and 5.5 mM glucose. High K+-KRB solution was prepared by replacing NaCl with KCl. Buffers containing intermediate K+ concentrations were prepared by mixing volumes of KRB and K+-KRB. Acetylcholine (ACh), phenylephrine (PHE), Nω-nitro-l-arginine methyl ester (L-NAME), sodium-nitroprusside (SNP) and indomethacin (INDO) were obtained from Sigma-Aldrich (Germany). All compounds were dissolved in KRB-buffer with the exception of INDO, which was dissolved in ethanol.
4
Artemisia asiatica Nakai Extract Modulation
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Flavonoids (luteolin, jaceosidin, and eupatilin) (Figure 1(a) ), indomethacin (Indo), N-nitro-L-arginine-methyl ester (L-NAME), polyethylenimine (PEI), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (a tetrazole) (MTT), sodium dodecyl sulfate (SDS), dimethyl sulfoxide (DMSO), pam3CSK, and lipopolysaccharide (LPS, E. coli 0111:B4) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). An ethanolic extract of Artemisia asiatica Nakai (Aa-EE) was used as reported previously [3 (link)]. Piceatannol (Picea) and PP2 were obtained from Calbiochem (La Jolla, CA, USA). The enzyme immune assay (EIA) kits that were used to determine PGE2 levels were purchased from Amersham (Little Chalfont, Buckinghamshire, UK). Fetal bovine serum (FBS), penicillin, streptomycin, TRIzol Reagent, and RPMI1640 were purchased from GIBCO (Grand Island, NY, USA). RAW264.7 and HEK293 cells were acquired from ATCC (Rockville, MD, USA). The rest of chemical reagents used in this study were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). Phosphospecific or total-protein antibodies raised against p65, p50, c-Fos, c-Jun, inhibitor of κBα (IκBα), Src, spleen tyrosine kinase (Syk), p85, ERK, JNK, p38, lamin A/C, and β-actin were obtained from Cell Signaling Technology (Beverly, MA, USA).
5
Pharmacological Modulation of Cellular Processes
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Acetylcholine (ACh, #A6625), indomethacin (indo, #I8280), PEG-catalase (cat, #C4963), sodium nitroprusside (SNP, #71778), SP600125 (#S5567), tamoxifen (#T5648) and TCS401 (#SML2140) were purchased from Sigma Aldrich (St Louis, MO, US). l -NAME (N(γ)-nitro-l -arginine methyl ester, #06-651-00), phenylephrine (#18-605-133), SB203580 (#12-021-0), streptozotocin (#57-220-1), thapsigargin (Thapsi, #50-464-293), Tauroursodeoxycholic Acid (TUDCA, #NC1266953), and tunicamycin (Tunica, #35-161-0) were purchased from Fisher Scientific (Waltham, MA, US). Tempol (#ALX-430-081-G001) was purchased from Enzo life sciences (Farmingdale, NY, YS).
6
Preparation of Krebs-Ringer Bicarbonate Buffer
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Krebs Ringer bicarbonate-buffered salt solution (KRB) contained (in mM): 118.5 NaCl, 4.7 KCl, 2.5 CaCl2, 1.2 MgSO4, 1.2 KH2PO4, 25.0 NaHCO3 and 5.5 glucose. The KRB solution was continuously aerated with 95% O2/5% CO2 and maintained at 37°C. Indomethacin (INDO; Sigma Aldrich, Zwijndrecht, NL) was dissolved in ethanol. Acetylcholine (ACh), noradrenaline (NA), phenylephrine (PHE), Nω-nitro-arginine methyl ester (L-NAME) and sodium nitroprusside (SNP; all Sigma Aldrich) were dissolved in KRB solution. High K+-KRB solution was prepared by replacing NaCl with KCl. Buffers containing intermediate K+ concentrations were prepared by mixing the appropriate volumes of KRB and K+-KRB.
7
Isolation and Characterization of ARR
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ARR (purity >95%) was isolated from Loranthus tanakae Franch. & Sav. in our laboratory, as previously described (21 (link)). The structure of ARR was elucidated by nuclear magnetic resonance. LPS and indomethacin (Indo) were purchased from Sigma-Aldrich; Merck KGaA. DMEM, FBS and penicillin-streptomycin were purchased from Gibco; Thermo Fisher Scientific, Inc. Primary rabbit monoclonal antibodies, including NF-κB p65 (cat. no. ab16502), phosphorylated-(p)-NF-κB-p65 (cat. no. ab76302), Nrf2 (cat. no. ab92946), NQO1 (cat. no. ab80588), HO-1 (cat. no. ab13243), TRAF6 (cat. no. ab137452), β-actin (cat. no. ab8227) and Histone H3 (cat. no. ab1791) were purchased from Abcam and the NLRC3 antibody (cat. no. DF13411) was obtained from Affinity Biosciences. The nitric oxide (NO) colorimetric kit (cat. no. E-BC-K035-M) and the cytokine mouse ELISA kits specific for prostaglandin E2 (PGE2; cat. no. E-EL-0034c), IL-6 (cat. no. E-EL-M0044c) and IL-1β (cat. no. E-EL-M0037c) were purchased from Elabscience.
8
Modulating Immune Responses with Inhibitors
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The inducible nitric oxide synthase (iNOS) inhibitor NG-methyl-L-arginine acetate salt (L-NMMA) and the cyclooxygenase (COX)-1/COX-2 inhibitor indomethacin (Indo) were purchased from Sigma (St. Louis, MO). An H-2b-restricted ovalbumin (OVA) class II epitope (323–339) was purchased from AnaSpec (Fremont, CA). TLR ligands, CpG1826 and CL097, were purchased from Invivogen (San Diego, CA). All the fluorescent-labeled antibodies were purchased from eBioscience (San Diego, CA). Recombinant mouse IL-2 was purchased from R & D systems (Minneapolis, MN) and neutralizing anti-IL-2 mAb (clone:JES6-1A12) was from Biolegend (San Diego, CA).
9
Anti-inflammatory Properties of SBKT Extract
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SBKT, isolated from the pulp, was obtained from Food and Herbal Park, Patanjali Ayurveda Limited, Haridwar, India. Culture media RPMI-1640, fetal bovine serum, and antibiotic/anti-mycotic mixture were obtained from Gibco. Cytokines interleukin 1-beta (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) ELISA kits were purchased from BD Biosciences. LPS, TPA, λ-Carrageenan, indomethacin (INDO), and dexamethasone (DEXA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Hematoxylin, potassium aluminium sulfate dodecahydrate, and mercury (II) oxide red were purchased from Merck India Pvt. Ltd, Mumbai, India. Eosin yellow and ferric chloride were purchased from Hi-Media Laboratories, Mumbai, India. All the other chemicals and reagents purchased for the study were of the highest commercial grade.
10
Vascular Smooth Muscle Pharmacology
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Acetylcholine chloride (ACh), Nω-Nitro-l -arginine methyl ester (l -NAME), phenylephrine (PE), tetraethylammonium (TEA), glibenclamide (Gliben), 1H-[1,2,4] oxadiazolo [4,3-α] quinoxaline-1-one (ODQ), and indomethacin (Indo) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). All drug solutions were dissolved in a solution containing NaCl 9 g/L, NaH2PO4 0.19 g/L, and ascorbic acid 0.03 g/L.
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