The agents were freshly prepared before the treatment experiments in the growth medium. The medium with a single agent was prepared as follows: 1) docetaxel (AbCam; Cambridge, United Kingdom) at final concentrations of 0.25 nM, 0.5, 1, 2, 4 nM, 8 and 16 nM; 2) JQ1 (Cayman Chemical, United States) at final concentrations of 8, 16, 32, 64, 128, and 256 nM. The medium of the mixture was prepared as follows: 1) 1 nM docetaxel combined with 128 nM JQ1; 2) 2 nM docetaxel combined with 128 nM JQ1). Culture medium without any agent was used as the negative control. The prepared medium was gently added to the cells (1 ml/well for 24 well plate, 200 ul/well for 96-well plate), and was changed every 3 days. The growth of cells or spheroids was observed using a TCS SPE confocal system microscope (Leica, Germany).
Docetaxel
Manufactured by Cayman Chemical
Sourced in United States
Docetaxel is a cytotoxic agent that functions as a microtubule inhibitor. It is a semi-synthetic taxane derivative used in the treatment of various types of cancer.
Automatically generated - may contain errors
Lab products found in correlation
Paclitaxel, by Cayman Chemical (2 mentions)
Vinblastine, by Cayman Chemical (2 mentions)
Etoposide, by Cayman Chemical (2 mentions)
Celltiter glo, by Promega (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
4 hydroxytamoxifen, by Cayman Chemical (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Docetaxel, by Abcam (1 mentions)
Hspa8, by Cell Signaling Technology (1 mentions)
Anxa2, by Cell Signaling Technology (1 mentions)
Succinate assay kit, by Abcam (1 mentions)
Hyou1, by Cell Signaling Technology (1 mentions)
Hsp90ab1, by Cell Signaling Technology (1 mentions)
Ki67 antibody, by Abcam (1 mentions)
β actin antibody, by Merck Group (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Oxaliplatin, by Merck Group (1 mentions)
Caffeic acid phenethyl ester, by Merck Group (1 mentions)
Fulvestrant, by Cayman Chemical (1 mentions)
15 protocols using docetaxel
1
Combination Therapy Evaluation: Docetaxel and JQ1
2
Establishing Hormone-Resistant Breast Cancer
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The triple-negative MDA-MB-231 and hormone-dependent MCF7 breast cancer cell lines were purchased from ATCC collection. The cells were maintained in a standard DMEM medium (Gibco) supplemented with 10% fetal bovine serum (FBS, HyClone) at 37 °C, 5% CO2 and 80%-85% humidity (NuAire CO2 incubator).
4-hydroxytamoxifen and docetaxel were purchased from Cayman Chemical Company; drug solutions were stored at -70 °C. MCF7/HT cell line was obtained by long-term (for 12 months) cultivation of parental MCF7 cells with antiestrogen 4-hydroxytamoxifen at a concentration of 5 μM. The verification of acquired hormone resistance in MCF7/HT was done by the MTT test.
4-hydroxytamoxifen and docetaxel were purchased from Cayman Chemical Company; drug solutions were stored at -70 °C. MCF7/HT cell line was obtained by long-term (for 12 months) cultivation of parental MCF7 cells with antiestrogen 4-hydroxytamoxifen at a concentration of 5 μM. The verification of acquired hormone resistance in MCF7/HT was done by the MTT test.
3
Dose-Dependent Cytotoxicity Assay
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Docetaxel (Cayman Chemical Company) was prepared as 5 mg/mL in DMSO and doxorubicin (AdooQ Bioscience) as 25 mg/mL in DMSO; both were then frozen into aliquots. Once thawed, Docetaxel and doxorubicin stock solutions were diluted in culture medium to give final working concentrations. Docetaxel dose-response analysis for both MDA-MB-231 and MCF-7 cells involved imaging eight wells treated with the following concentrations of Docetaxel: 0 nM, 1 nM, 3 nM, 10 nM, 30 nM, 100 nM, 300 nM, 1 μM, with additional concentrations 3 μM, 10 μM and 30 μM imaged for MDA-MB-231 cells. Doxorubicin dose-response analysis for MDA-MB-231 cells involved imaging eight wells treated with the following concentrations of doxorubicin: 0 nM, 10 nM, 30 nM, 100 nM, 300 nM, 1 μM, 3 μM, 10 μM.
Medium was removed from wells selected to receive treatment 30 mins prior to image acquisition, and 500 μL of desired drug concentration was added to each well. Control wells received a medium change and were treated with DMSO vehicle on the day of imaging to maintain consistent DMSO concentration throughout.
Medium was removed from wells selected to receive treatment 30 mins prior to image acquisition, and 500 μL of desired drug concentration was added to each well. Control wells received a medium change and were treated with DMSO vehicle on the day of imaging to maintain consistent DMSO concentration throughout.
4
Characterization of Prostate Cancer Cell Lines
5
Cytotoxic Drug Treatment in CRC and PCa
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Oxaliplatin (Sigma Aldrich, St Louis, MO, USA) or docetaxel (Cayman Chemical, Ann Arbor, MI, USA) was used at the indicated concentrations to treat CRC or PCa cells, respectively.
6
Reagents for In Vitro Experiments
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Caffeic acid phenethyl ester (CAPE) was purchased from Sigma-Aldrich (St. Louis, MO, U.S.A.), while docetaxel was purchased from Cayman Chemicals (Ann Arbor, MI, U.S.A.).
7
Cytotoxicity Assay of Chemotherapies
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2.5 × 103 A594 or H460 cells in 96-well opaque plates and allowed to proliferate for 24 hours before treating with 10-fold dilutions of docetaxel (Cayman Chemical, Ann Arbor, MI, USA) #11637), paclitaxel (Cayman Chemical #10461), etoposide (Cayman Chemical #12092), vinblastine (Cayman Chemical #11762), and gemcitabine (Duke Pharmacy Store Room, Durham, NC, USA). Cells were treated with chemotherapy reagents for 48 hours, and cell viability was assessed using CellTiter-Glo (Promega, Madison, WI, USA) and normalized to an untreated control.
8
Evaluating Efficacy of Estrogen Modulators
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27-Hydroxycholesterol was purchased from Santa Cruz Biotechnologies (Dallas, TX), docetaxel, 4-[2-phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5-a]pyrimidin-3-yl]phenol (PHTPP) and fulvestrant from Cayman Chemicals (Ann Arbor, MI) and β-estradiol from Sigma-Aldrich (St. Louis, MO). All cell culture reagents, with the exception of fetal bovine serum (FBS) (Atlanta Biologicals; Flowery Branch, GA) were from Invitrogen (Carlsbad, CA). Human RWPE-1, LNCaP and PC3 cells were purchased from ATCC (Manassas, VA).
9
Vascular Function Assay with NACT Drugs
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Arterial rings from no-NACT patients were incubated with the individual components of NACT: docetaxel, doxorubicin, cyclophosphamide, 4-hydroperoxycyclophosphamide (all from Cayman Chemicals), and vehicle (solvent) as a control. The drug concentrations used were 10 nM or 100 nM. Arteries were incubated in a buffer containing HBSS (Gibco, Thermo Fisher Scientific), 20 mM HEPES (Gibco, Thermo Fisher Scientific), and gentamicin with glutamine in an incubator at 37°C in a humidified atmosphere for 24 hours (52 (link)). After incubation, vascular function was measured, or rings were placed in RNAlater (Ambion) for gene expression and RNA-Seq studies.
10
Chemotherapeutic Cytotoxicity Assay Optimization
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Cell lines were seeded in 96-well plates, at a concentration of 2000–4000 cells per well (optimized to achieve 100% confluency at 72 hours in untreated/control wells), and allowed to adhere overnight (12–16 hours). Medical grade carboplatin, mitoxantrone, and vinblastine were used in these studies. Docetaxel was purchased (Cayman Chemical, 11,637) and solubilized in DMSO. Treatments ensued for 72 hours with each chemotherapeutic prior to completing SRB assays as previously described [27 (link)]. Several smaller cohort cytotoxicity experiments (evaluating a few cell lines at a time) were performed to optimize seeding densities and drug concentration ranges before completing the full cell line cytotoxicity experiment. Based on these findings, the following drug concentrations were selected for the full cell line cytotoxicity experiment: vinblastine (0.01pM-100 μM), Docetaxel (10pM-1 μM), mitoxantrone (0.3 nM-30 μM), and carboplatin (0.39 μM–400 μM). Each drug concentration was performed in four replicate wells. Magnitude of chemotherapy-induced cytotoxicity was normalized against untreated control (carboplatin, mitoxantrone, and vinblastine) or DMSO control (Docetaxel). Results were graphed using ggplot2 (version 3.3.5) in RStudio (RStudio 2021, Boston, MA), and data grouped based upon cancer of origin (iUC versus PCA).
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