Libraries used for high-throughput drug screening are described in Figure 1 . Other compounds used include the HDAC inhibitors SAHA, HNHA, LBH-589, Scriptaid, MS-275, Givinostat, PDX101, LAQ-824 and MGCD0103 (all from Selleck), the PP2A inhibitors Cantharidin and NorCantharidin, the topoisomerase inhibitors Camptothecin and Topotecan (all from Sigma), the HMG CoA reductase inhibitors Cerivastatin (Sigma) and Itavastatin (Sequoia Research Products Ltd. UK), and the PI3K/mTOR inhibitors BEZ-235 and BKM-120 (both from Active Biochem). All inhibitors were dissolved in DMSO to 20 mM, and further diluted to an appropriate final concentration in culture medium at the time of use.
Cerivastatin
Manufactured by Merck Group
Sourced in United Kingdom
Cerivastatin is a laboratory equipment product manufactured by the Merck Group. It is a statin medication used to lower cholesterol levels in the body. Cerivastatin functions by inhibiting the enzyme HMG-CoA reductase, which is responsible for the production of cholesterol in the liver.
Automatically generated - may contain errors
Lab products found in correlation
Simvastatin, by Merck Group (4 mentions)
Fluvastatin, by Merck Group (3 mentions)
Squalene, by Merck Group (2 mentions)
Atorvastatin, by Merck Group (2 mentions)
Camptothecin, by Merck Group (1 mentions)
Norcantharidin, by Merck Group (1 mentions)
Lbh589, by Selleck Chemicals (1 mentions)
Ms 275, by Selleck Chemicals (1 mentions)
Givinostat, by Selleck Chemicals (1 mentions)
Cantharidin, by Merck Group (1 mentions)
Topotecan, by Merck Group (1 mentions)
Bkm120, by Active Motif (1 mentions)
Scriptaid, by Selleck Chemicals (1 mentions)
Mgcd0103, by Selleck Chemicals (1 mentions)
Bez 235, by Active Motif (1 mentions)
Geneticin g418, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Hygromycin, by Merck Group (1 mentions)
12 protocols using cerivastatin
1
High-throughput Drug Screening Compounds
2
Cell Culture and Molecular Biology Protocols
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Reagents for cell culture, buffer preparation and molecular biology were purchased from Merck/Sigma-Aldrich, (St. Louis, MO, USA). Simvastatin, cerivastatin, propidium iodide, puromycin, hygromycin, doxycycline and DAPI were purchased from Merck/Sigma-Aldrich. Geneticin (G-418) was from Gibco (Fisher Scientific, Loughborough, UK). XAV939 was obtained from Selleckchem (Houston, TX, USA). Human His-tagged recombinant TRAIL was produced as described [34 (link)].
3
Cell Line Treatments with Statins, CDK4/6 Inhibitors, and MEK Inhibitors
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MDA-MB-231 and MDA-MB-157 cell lines were obtained from ATCC. All cell lines were grown in DMEM with 4.5 g/L glucose (Lonza) and L-glutamine, supplemented with 10% fetal bovine serum (Euroclone), 100 U/ml penicillin and 100 U/ml streptomycin (Lonza) at 37°C in a humidified atmosphere of 5% CO2.
Cerivastatin (SML0005, Sigma Aldrich) and DL-mevalonic acid 5-phosphate (79849, Sigma Aldrich,) were prepared in dimethyl sulfoxide at a 10mmol/L concentration and 0.5mol/L, respectively.
Palbociclib (PD0332991, PZ 0199, Sigma Aldrich) and abemaciclib (LY2835219, HY-16297, MedChem express) were dissolved in H2O at a 1mmol/L concentration. RRD-251 hydrochloride (R7532, Sigma Aldrich) was resuspended in dimethyl sulfoxide at a 50mmol/L concentration.
For Cerivastatin treatment, cells were incubated with DMSO or 1μM Cerivastatin alone or with 0.5mmol mevalonic acid (MVA) for 24 hours (MDA-MB-231) or 72 hours (MDA-MB-157).
For Palbociclib treatment, cells were treated with 0.5μM (MDA-MB-231) or 1μM (MDA-MB-157) PD0332991 for 24 hours. Treatment with abemaciclib was performed incubating cells with 0.5μM LY2835219 for 24 hours. For RRD-251 treatment cells were incubated with DMSO or 100μM RRD-251 for 24 hours.
Cerivastatin (SML0005, Sigma Aldrich) and DL-mevalonic acid 5-phosphate (79849, Sigma Aldrich,) were prepared in dimethyl sulfoxide at a 10mmol/L concentration and 0.5mol/L, respectively.
Palbociclib (PD0332991, PZ 0199, Sigma Aldrich) and abemaciclib (LY2835219, HY-16297, MedChem express) were dissolved in H2O at a 1mmol/L concentration. RRD-251 hydrochloride (R7532, Sigma Aldrich) was resuspended in dimethyl sulfoxide at a 50mmol/L concentration.
For Cerivastatin treatment, cells were incubated with DMSO or 1μM Cerivastatin alone or with 0.5mmol mevalonic acid (MVA) for 24 hours (MDA-MB-231) or 72 hours (MDA-MB-157).
For Palbociclib treatment, cells were treated with 0.5μM (MDA-MB-231) or 1μM (MDA-MB-157) PD0332991 for 24 hours. Treatment with abemaciclib was performed incubating cells with 0.5μM LY2835219 for 24 hours. For RRD-251 treatment cells were incubated with DMSO or 100μM RRD-251 for 24 hours.
4
Modulating Yap Signaling in Cardiac Reprogramming
5
Evaluating Statin Effects on Neurons
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Atorvastatin, lovastatin, and pitavastatin were obtained from Selleckchem Inc. Cerivastatin was obtained from Sigma. All statins were > 98% pure based on HPLC data and reconstituted in dimethyl sulfoxide (DMSO, Sigma). Mevalonate (Sigma) was prepared in sterile‐filtered purified water. Human primary neuron cells were obtained from ScienCell Inc. Neuron cell line SH‐SY5Y (human) and PC12 (rat) were obtained from American Type Culture Collection. All cells were cultured using the same medium as described in our previous study.14 All cells were examined for mycoplasma using a MycoAlert Mycoplasma Detection kit (Lonza) prior to experiments. Conduct of these experiments were not blinded for each operator as measurements were not prone to bias. Deductions from each experimental setup were derived from a collective analysis of all datasets and not just based on a single condition.
6
High-Throughput Screening of Diverse Chemical Libraries
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The screen consisted of 5 libraries: Prestwick chemical library (1280 compounds), Library of Pharmacologically Active Compounds (LOPAC)(1280 compounds), FDA/International drug library (1280 compounds), NIH clinical collection (446 compounds), and NCI oncology library (89 compounds). In total, 4,375 total drugs were tested. For confirmatory studies, all statins were obtained as dry powder from Selleckchem with the exception of cerivastatin which was purchased from Sigma. The purity of all compounds was verified by LC-MS analysis with all found to be >95% pure.
7
Evaluation of FDA-Approved Drugs for Novel Activities
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Crizotinib, ceritinib, and lorlatinib were purchased from Selleck Chemical (Houston, TX). The library of FDA‐approved drugs (Screen‐Well FDA‐Approved Drug Library, 640 chemical compounds dissolved at 10 mM in dimethyl sulfoxide) was obtained from Enzo Life Sciences (Plymouth Meeting, PA). The following compounds were purchased from Sigma Chemical Co. (St. Louis, MO): cerivastatin, simvastatin, fluvastatin, atorvastatin, GGPP, FPP, squalene, GGTI‐298, FTI‐277, and verteporfin. Drug preparation and use of all reagents were conducted according to manufacturer's instructions.
8
Cerivastatin Effects on Neurosphere Proliferation
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Neurospheres were dissociated and seeded in triplicate for each biological replicate (isolated from individual embryos) in proliferation media at a density of 10,000 cells/well in a 96-well plate (#CC7682-7596; USA Scientific) coated with laminin (10 µg/ml) and allowed to recover for 18–22 h. To visualize nuclei, 100 µl proliferation media containing 250 nM siR-DNA Kit (#CY-SC007; Cytoskeleton, Inc.) was added per well. Cerivastatin (#SML0005; Sigma-Aldrich) was added at 0, 20, 50, and 100 nM. Live cells were imaged with a 10× phase objective with both phase contrast and fluorescence (using filter cube Ex 565–605 nm/Em 625–705 nm) at four to five positions per well every 2 h in the IncuCyte S3 Live-Cell Analysis System for 60 h. Fold proliferation was determined by analysis with Sartorius software of the number of nuclei at each time point in each image and normalized to the number of nuclei at time point 0 for each image. Data presented are from three wells for each embryo. A minimum of three embryos for WT and for KO were used per experiment.
9
Investigating Cancer Cell Modulation by Retinoic Acid and Kinase Inhibitors
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MDA-MB-231 and MDA-MB-468 cells, obtained from the American Type Culture Collection (ATCC) were maintained in Dulbecco’s modified Eagle medium/Ham’s F12 (1:1) supplemented with 10% fetal bovine serum (FBS) and 2 mM glutamine. Human umbilical vein endothelial cells (HUVEC) were obtained from Clonetics (cc-2517) and cultured following the provider’s recommendations and media. Cells were maintained in a 5% CO2 atmosphere at 37 °C. All cell lines were used at low passage (<20) and regularly tested against mycoplasma.
All-trans-retinoic acid (RA) (Sigma) was dissolved in ethanol (stock solution 2.5 mM). The reagent was diluted to its final concentration (5 μM) using cell culture medium. The selective Src family-tyrosine kinase inhibitor, 4-amino-5- (4-chlorophenyl)-7-(t-butyl) pyrazolo (3,4-d) pyrimidine (PP2) (Calbiochem, San Diego, CA)35 (link) suspended in 1% dimethyl sulfoxide (DMSO) was added to the culture medium at a concentration of 10 μM. Cerivastatin (Sigma) was dissolved to a final concentration of 1 μM in the culture medium. Cells were treated for 48 hrs with either ethanol, DMSO, 10 μM PP2, 1 µM Cerivastatin or 1 µM Cerivastatin plus RA (5 μM).
All-trans-retinoic acid (RA) (Sigma) was dissolved in ethanol (stock solution 2.5 mM). The reagent was diluted to its final concentration (5 μM) using cell culture medium. The selective Src family-tyrosine kinase inhibitor, 4-amino-5- (4-chlorophenyl)-7-(t-butyl) pyrazolo (3,4-d) pyrimidine (PP2) (Calbiochem, San Diego, CA)35 (link) suspended in 1% dimethyl sulfoxide (DMSO) was added to the culture medium at a concentration of 10 μM. Cerivastatin (Sigma) was dissolved to a final concentration of 1 μM in the culture medium. Cells were treated for 48 hrs with either ethanol, DMSO, 10 μM PP2, 1 µM Cerivastatin or 1 µM Cerivastatin plus RA (5 μM).
10
Lipid Biosynthesis Pathway Regulation
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LPS was obtained from Invitrogen (Carlsbad, CA). Cerivastatin, simvastatin, fluvastatin, pravastatin, cytochalasin D, and squalene were from Sigma-Aldrich (St. Louis, MO). GG-PP, Ro 48-8071, and YM-53601 were purchased from Cayman Chemical (Ann Arbor, MI). GGTI 298 was purchased from R&D systems (Minneapolis, MN). Mevalonate was from Santa Cruz (Dallas, TX).
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