Fenofibrate and bezafibrate was obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). GW6471, ANIT, formic acid, chlorpropamide, lauroylcarnitine (12:0-carnitine), myristoylcarnitine (14:0-carnitine), palmitoylcarnitine (16:0-carnitine), stearoylcarnitine (18:0-carnitine), CA, UDCA, HDCA, CDCA, DCA, TCA, GCA, THDCA, TCDCA, TDCA, TLCA, glycochendeoxycholic acid (GCDCA), glycodeoxycholic acid (GDCA), dehydrocholic acid (DHCA), and glycolithocholic acid (GLCA) were purchased from Sigma-Aldrich (St. Louis, Missouri, USA). Hyocholic acid (HCA), TβMCA, and ωMCA were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). TUDCA and LCA was purchased from Medchemexpress (Monmouth Junction, NJ, USA). TαMCA and βMCA were purchased from Steraloids (Newport, RI). All solvents and organic reagents were of the highest grade commercially available.
Tcdca
Manufactured by Merck Group
Sourced in United States, Germany
TCDCA is a laboratory reagent used in various analytical and research applications. It is a bile acid derivative that serves as a chemical standard or reference material for analytical techniques. The core function of TCDCA is to provide a consistent and reliable reference point for the identification and quantification of similar compounds in samples.
Automatically generated - may contain errors
Lab products found in correlation
Gcdca, by Merck Group (11 mentions)
T β mca, by Steraloids (4 mentions)
T udca, by Steraloids (4 mentions)
T hdca, by Steraloids (4 mentions)
G lca, by Steraloids (3 mentions)
G udca, by Steraloids (3 mentions)
G dca, by Steraloids (3 mentions)
Tudca, by Merck Group (3 mentions)
Hyodeoxycholic acid, by Merck Group (3 mentions)
Tβmca, by Merck Group (3 mentions)
Tαmca, by Steraloids (2 mentions)
Formic acid, by Merck Group (2 mentions)
T lca, by Steraloids (2 mentions)
Deoxycholic acid, by Merck Group (2 mentions)
Cholic acid, by Merck Group (2 mentions)
Tudca, by MedChemExpress (1 mentions)
Chlorpropamide, by Merck Group (1 mentions)
Lauroylcarnitine, by Merck Group (1 mentions)
Myristoylcarnitine, by Merck Group (1 mentions)
Palmitoylcarnitine, by Merck Group (1 mentions)
20 protocols using tcdca
1
Analytical Standards for Bile Acids
2
Inflammatory Bowel Disease Biomarker Analysis
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OEA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); DSS (molecular weight=36,000–50,000 kDa) was obtained from MP Biochemical (Santa Ana, CA). MPA O-glucuronide and naloxone 3-β-D-glucuronide were obtained from Toronto Research Chemicals Inc. (ON, Canada). β-MCA, UDCA, HDCA, G-LCA, G-UDCA, G-DCA, T-β-MCA, T-UDCA, T-HDCA and T-LCA were purchased from Steraloids Inc. (Newport, RI). α-MCA, CA, DCA, CDCA, lithocholic acid (LCA), G-CDCA, G-CA, T-CDCA, T-DCA, T-CA, dehydrocholic acid and propranolol were purchased from Sigma-Aldrich (St Louis, MO). The anti-UGT1A, anti-FXR, anti-FGF15 and anti-CYP7A1 antibodies were purchased from Santa Cruz Biotechnology. The anti-PPARα antibody was from Abcam (Cambridge, UK). The anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from SunShine Biotechnology (Nanjing, China). The secondary antibodies and recombinant human FGF19 was obtained from Bioworld Technology (St Louis Park, MN). Other reagents, unless mentioned, were obtained from Sigma-Aldrich. RNA extracts of colon biopsies from 8 healthy humans and 13 ulcerative colitis and Crohn’s disease patients were previously described51 (link). The ulcerative colitis and Crohn’s disease samples were collected from consented patients and approved by the University of Michigan IRB committee (approval number HUM00042210).
3
Bile Acid Profiling Protocol
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Profiling of the principal human bile acids was performed as described previously10 (link). Twelve bile acid standards were used and resuspended in methanol to a concentration of 20 mM; CA, CDCA, DCA, LCA, UDCA, GDCA, TCDCA, TDCA, TCA, GCA, and TLCA were purchased from Sigma-Aldrich (Buchs, Switzerland). TUDCA was purchased from Calbiochem (Darmstadt, Germany). The twelve stock solutions were then pooled together to a concentration of 100 μM in methanol. All chemicals used were LC-MS grade. Duplicate samples were included in the analysis to ensure reproducibility and all analyses were performed blinded to patient data.
4
Quantification of Bile Acids Metabolites
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ANIT, carbamazepine, mycophenolic acid, and reference standards for 18 BAs, including CA, glycocholic acid, TCA, CDCA, glycochenodeoxycholic acid, TCDCA, deoxycholic acid, glycodeoxycholic acid, taurodeoxycholic acid, UDCA, glycoursodeoxycholic acid, TUDCA, LCA, glycolithocholic acid, taurolithocholic acid, hyodeoxycholic acid, glycohyodeoxycholic acid, and taurohyodeoxycholic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Chromatography-grade acetonitrile was purchased from Merck (Darmstadt, Germany). Water was purified using a MilliQ water system (Millipore, Bedford, MA, USA). Chromatography-grade acetic acid, formic acid, and methanol were provided by Tedia Company (Fairfield, OH, USA).
5
Isolation and Characterization of Mouse Hepatic Stellate Cells
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Isolation of primary mHSC was performed via pronase–collagenase perfusion followed by density gradient centrifugation in 13.2% Nycodenz (Axis-Shield PoC, Oslo, Norway) [33 (link)]. Purity of preparation was assessed by confirmation of vitamin A autofluorescence.
Isolated HSCs were allowed to attach for 2 h and were then stimulated with bile salts CDCA, GCDCA, TCDCA, and UDCA (Sigma-Aldrich, Darmstadt, Germany) for the time periods indicated, in the absence or presence of AG1478 (Sigma-Aldrich, Darmstadt, Germany) and PD98059 (Cayman, Ann Arbor, MI, USA). To quantify total DNA as a surrogate of cell number, HSCs were incubated with PicoGreen® (Invitrogen, Carlsbad, CA, USA) and fluorescence signals were detected with a CytoFluor 4000 system (PerSeptive Biosystems, Framingham, MA, USA). Proliferation of HSC was quantified using a BrdU-assay kit (Roche, Penzberg, Germany) according to the manufacturer’s instructions. To quantify total cell count, HSCs, seeded in Lab-Tek II Chamber Slides (Nunc, Rochester, NY, USA), were mounted on cover slides with Vectashield mounting medium including DAPI (Vector, Burlingame, CA, USA). Slides were scanned with a Pannoramic Midi Slide Scanner (3DHistech, Budapest, Hungary) and nucles count was performed with ImageJ2 software on the complete slide (0.7 cm2).
Isolated HSCs were allowed to attach for 2 h and were then stimulated with bile salts CDCA, GCDCA, TCDCA, and UDCA (Sigma-Aldrich, Darmstadt, Germany) for the time periods indicated, in the absence or presence of AG1478 (Sigma-Aldrich, Darmstadt, Germany) and PD98059 (Cayman, Ann Arbor, MI, USA). To quantify total DNA as a surrogate of cell number, HSCs were incubated with PicoGreen® (Invitrogen, Carlsbad, CA, USA) and fluorescence signals were detected with a CytoFluor 4000 system (PerSeptive Biosystems, Framingham, MA, USA). Proliferation of HSC was quantified using a BrdU-assay kit (Roche, Penzberg, Germany) according to the manufacturer’s instructions. To quantify total cell count, HSCs, seeded in Lab-Tek II Chamber Slides (Nunc, Rochester, NY, USA), were mounted on cover slides with Vectashield mounting medium including DAPI (Vector, Burlingame, CA, USA). Slides were scanned with a Pannoramic Midi Slide Scanner (3DHistech, Budapest, Hungary) and nucles count was performed with ImageJ2 software on the complete slide (0.7 cm2).
6
Comprehensive Bile Acid Profiling Protocol
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HPLC-grade methanol and acetonitrile were purchased from Fisher Scientific (Nepean, Ont., Canada). Ultrapure water was prepared by the Milli-Q Ultrapure water purification system (Millipore, Bedford, MA, USA). AOM was obtained from Sigma-Aldrich (St. Louis, MO, USA), and DSS (MW 36000-50000) was purchased from MP Biomedicals (Santa Ana, CA). Ammonium acetate, formic acid, glacial acetic acid and the other reagents (analytical grade) were purchased from Nanjing Chemical Factory (Nanjing, China). ‘Mouse Total Bile Acids Kit’ was purchased from Crystal Chem Inc (Downers Grove, United States). ‘MiniBEST Bacterial Genomic DNA Extraction Kit’ was from TaKaRa Bio Inc (Dalian, China). Anti-FXR, FGF15 and CYP7A1 antibodies were all purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and anti-GAPDH antibody was from Bioworld Technology (St. Louis Park, MN, USA).
CA, CDCA, DCA, and LCA, as well as their glycine [38 (link)] and taurine [45 (link)] conjugates, G-CA, G-CDCA, T-β-MCA, T-CA, T-CDCA, T-DCA, and internal standard dehydrocholic acid (dhCA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). α-MCA, UDCA, HDCA, G-LCA, G-UDCA, G-DCA, T-β-MCA, T-UDCA, T-HDCA and T-LCA were purchased from Steraloids, Inc. (Newport, Rhode Island, USA).
CA, CDCA, DCA, and LCA, as well as their glycine [38 (link)] and taurine [45 (link)] conjugates, G-CA, G-CDCA, T-β-MCA, T-CA, T-CDCA, T-DCA, and internal standard dehydrocholic acid (dhCA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). α-MCA, UDCA, HDCA, G-LCA, G-UDCA, G-DCA, T-β-MCA, T-UDCA, T-HDCA and T-LCA were purchased from Steraloids, Inc. (Newport, Rhode Island, USA).
7
Bile Acid Panel Profiling Protocol
8
Bile Acid Signaling Regulation
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Special reagents used in the study included dimethyl sulfoxide (DMSO) and various BAs of UDCA, GCA, TCDCA, TMCA, GCDCA, and TDCA (Sigma-Aldrich Chemical); [3H]-TCA (1 mCi/mL, PerkinElmer Life and Analytical Sciences)23 (link); human recombinant IL-6, IL-1β, TNFα, and CCL2 (Peprotech); and APTSTAT3-9R (Cat#S8197), an inhibitor of STAT3 phosphorylation (Selleck Chem).
9
Quantification of Bile Acids by HPLC
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SBA standards of lithocholic acid 3-sulfate (LCA-3S), glycolithocholic acid 3-sulfate (GLCA-3S) and taurolithocholic acid 3-sulfate (TLCA-3S) were purchased from Sigma–Aldrich (St. Louis, MO, USA). The authentic compounds of five kinds of unconjugated BAs, including cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), ursodeoxycholic acid (UDCA), and hyodeoxycholic acid (HDCA), and four kinds of glycine conjugated BAs including GCA, GCDCA, GDCA and GUDCA, and four kinds of taurine conjugated BAs including TCA, TCDCA, TDCA, and TUDCA were all obtained from Sigma–Aldrich (St. Louis, MO, USA). 5β-cholanicacid-3α, 6β, 7α-triol, as internal standard (IS), was purchased from Steraloids Chemical (Newport, Rhode Island, USA). Water was prepared by a MilliQ™ System (Millipore, Milford, MA, USA). HPLC-grade methanol (MeOH) and acetonitrile (ACN) were obtained from Merck KGaA (Merck KGaA, Darmstadt, Germany).
10
Comprehensive Bile Acid Quantification
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Deuterated-cholic acid (d4-CA), bile acids (CA, CDCA, LCA, TCA, UDCA, taurine, βMCA, TαMCA, TβMCA, TLCA, TCDCA), and formic acid were purchased from Sigma-Aldrich Chemicals Co. (St Louis, MO, USA). All other bile acid standards (GCDCA, GCA, GDCA, GHDCA, GLCA, GUDCA, HCA, HDCA, ILA, TDCA, THDCA, TUDCA) were purchased from Steraloids Inc. (Newport, RI, USA). Acetonitrile (ACN) and methanol (MeOH) of Optima LC-MS grade quality were purchased from Thermo Fisher Scientific (Auckland, New Zealand).
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