Citco

Manufactured by Merck Group

Sourced in United States

CITCO is a laboratory equipment product manufactured by Merck Group. It is designed for performing cell-based assays and other in vitro experiments. The core function of CITCO is to provide a controlled environment for cell culture and analysis.

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Lab products found in correlation

5 protocols using citco

Analytical Methods for Anticancer Compounds

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CITCO, CPA, doxorubicin, prednisone, vincristine, hexamethyl phosphoramide (HMP), chrysin, 1,4-Bis [2-(3,5-Dichloropyridyloxy)] benzene (TCPOBOP), semicarbazide, ammonium formate, methyl-tert butyl ether (MTBE), isopropyl alcohol (IPA) formic acid (FA) and corn oil were all purchased from Sigma Aldrich (St. Louis, MO, USA). 4-hydroxy cyclophosphamide (4-OH-CPA) was purchased from Toronto Research Chemicals (North York, ON, Canada). Optima LC/MS grade acetonitrile (ACN), methanol (MeOH), water (H₂O) were obtained from Fisher Scientific (Pittsburg, PA, USA). Sterile normal saline solution was obtained from Vedco (St. Joseph, MO, USA). DL5009 was synthesized at the University of Maryland School of Pharmacy as described previously [27 (link)].

Kurian R., Hedrich W., Mackowiak B., Li L, & Wang H. (2020). CITCO as an Adjuvant Facilitates CHOP-Based Lymphoma Treatment in hCAR-Transgenic Mice. Cells, 9(11), 2520.

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Ligand Screening for Nuclear Receptors

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Compound 1 (CITCO, (6-(4-chlorophenyl)imidazo [2,1-b][1,3]thiazole-5-carbaldehyde-O-(3,4-dichloro-benzyl)oxime)),
rifampicin (rif), TCPOBOP, and PK11195 were obtained from Sigma-Aldrich
(St. Louis, Missouri, United States, now Merck), which is now known
as Merck (Darmstadt, Germany). Phenobarbital (Luminal 200 mg/mL injection)
was manufactured by Desitin Pharma spol. s.r.o. (Prague, Czech Republic).
Ligands for nuclear receptors (GW3965, thyroxin, obeticholic acid,
dexamethasone, fenofibrate, GW501516, rosiglitazone, 3-methylcholanthrene,
calcitriol, testosterone, and estradiol) were purchased from Sigma-Aldrich
(now Merck). The prototype ligands were used at 100 nM (dexamethasone,
calcitriol), 1 μM (thyroxin), or 10 μM concentrations.
The compounds were dissolved in DMSO, and the final concentration
of DMSO in the entire reaction mixture or cultivation media was 0.1%.

Mejdrová I., Dušek J., Škach K., Stefela A., Skoda J., Chalupský K., Dohnalová K., Pavkova I., Kronenberger T., Rashidian A., Smutná L., Duchoslav V., Smutny T., Pávek P, & Nencka R. (2023). Discovery of Novel Human Constitutive Androstane Receptor Agonists with the Imidazo[1,2-a]pyridine Structure. Journal of Medicinal Chemistry, 66(4), 2422-2456.

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Visualizing CAR and YAP in CITCO-Treated HepG2 Cells

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According to previous publications²⁴ (link), HepG2 cells (ATCC, Cat# HB-8065, RRID: CVCL_0027) were cultured in DMEM containing 10% FBS and then treated with DMSO or 10 μmol/L CITCO (Sigma–Aldrich, Cat# C6240) for 48 h. After incubation, cells were fixed in 4% paraformaldehyde and 0.5% Triton X-100, respectively. Then, cells were incubated with rabbit polyclonal anti-CAR (Abcam, Cat# ab62590, RRID: AB_956175) and mouse monoclonal anti-YAP (R&D Systems, Cat# MAB8094, RRID: AB_2819224, MN, USA) overnight at 4 °C. Cells were stained with secondary antibodies including anti-mouse IgG Alexa Fluor 488 (Cell Signaling Technology, Cat# 4408, RRID: AB_10694704) and anti-rabbit IgG Alexa Fluor 647 (Cell Signaling Technology, Cat# 4414, RRID: AB_10693544). Images were acquired using a confocal microscope (Olympus FV3000, Tokyo, Japan).

Gao Y., Fan S., Li H., Jiang Y., Yao X., Zhu S., Yang X., Wang R., Tian J., Gonzalez F.J., Huang M, & Bi H. (2020). Constitutive androstane receptor induced-hepatomegaly and liver regeneration is partially via yes-associated protein activation. Acta Pharmaceutica Sinica. B, 11(3), 727-737.

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Nuclear Receptor Modulator Assay

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Paclitaxel, CITCO, TCPOBOP, androstenol and MTT were obtained from Sigma-Aldrich (St. Louis, MO, USA). Media and reagents for cell culture were acquired from Invitrogen (Carlsbad, CA, USA). Trizol, oligoDT primers, Superscript II enzyme and the Power SYBR Green mastermix were from Life Technologies (Grand Island, NY, USA). Other reagents were of analytical grade. The cell lines used in this experiment were the mouse cell line E9 [21] (link) and the human cell lines A549, H2023, H460, H2030, H1792 and H23 [22] (link). All these cell lines were a gift from Dr. Lucy M. Anderson from the Laboratory of Comparative Carcinogenesis at the Frederick National Laboratory for Cancer Research (United States of America).

Fukumasu H., Rochetti A.L., Pires P.R., Silva E.R., Mesquita L.G., Strefezzi R.F., De Carvalho D.D, & Dagli M.L. (2014). Constitutive Androstane Receptor Ligands Modulate the Anti-Tumor Efficacy of Paclitaxel in Non-Small Cell Lung Cancer Cells. PLoS ONE, 9(6), e99484.

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Screening and Evaluation of Compounds for Metabolic Regulation

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PB and CITCO were obtained from Sigma-Aldrich (St. Louis, MO). All 144 compounds (designated as UM050-UM200) were obtained from Specs (Delft, The Netherlands). The Dual-Luciferase Reporter Assay System was purchased through Promega (Madison, WI). Antibodies against CYP2B6, PEPCK, G6Pase, fatty acid synthase (FAS), and SREBP-1c were purchased from Santa Cruz (Dallas, TX). The acetyl-coA carboxylase 1 (ACC-α) and stearoyl-coA desaturase-1 (SCD-1) antibodies were acquired from Cell Signaling (Danvers, MA). β-actin antibody was from Sigma-Aldrich. Matrigel, insulin, and ITS⁺ (insulin/transferrin/selenium) were from BD Biosciences (Bedford, MA). The glucose and triglyceride assay kits were from Sigma-Aldrich. All other cell culture reagents were purchased from Life Technologies (Grand Island, NY) or Sigma-Aldrich.

Lynch C., Pan Y., Li L., Heyward S., Moeller T., Swaan P.W, & Wang H. (2014). Activation of the Constitutive Androstane Receptor Inhibits Gluconeogenesis without Affecting Lipogenesis or Fatty Acid Synthesis in Human Hepatocytes. Toxicology and applied pharmacology, 279(1), 33-42.

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