LNCaP cell line was obtained from ATCC, the v-Src PEC and the NeuT-PEC cell lines were previously described (18 (link)). Original cells were expanded and stored in the liquid nitrogen at early passage. During the experiments, the morphology of all cell lines was checked under phase contrast microscope routinely. For LNCaP cell line, proliferation and AR abundance in response to DHT stimulation were tested by MTT assay and Western-blot. For v-Src-PEC and NeuT-PEC cell lines, the proliferation in response to Src kinase inhibitor or NeuT inhibitor was tested. V-Src or NeuT expression in these cells was checked by Western-blot for verification. All of the newly revived cells were treated with BM-cyclins (Roche) and the mycoplasma contamination was determined with Hoechst 33258 staining under high magnification fluorescent microscope routinely. DNA transfection and luciferase assays were performed as previously described (1 (link),18 (link)). The CBF-Luc and -3,400 cyclin D1-Luc reporter plasmids were previously described (19 (link),20 (link)). The Src kinase inhibitor PP1 (4-amino-5-(4-methylphenyl)-7-(t-butel)pyrazolo-d-3-4-pyrimidine (Calbio Chem) and Dasatinib (BMS-354825, Selleckchem), the CDK inhibitor Abemaciclib (MedChem Express), Palbociclib (Sigma-Aldrich), Ribociclib (Selleckchem) and the EGFR inhibitor Canertinib (Selleckchem) were used at the indicated doses.
Bm cyclin
Manufactured by Roche
Sourced in Germany
The BM-Cyclin is a versatile laboratory equipment designed for use in various research and diagnostic applications. It serves as a tool for the analysis and detection of specific biomolecules, such as proteins and nucleic acids. The core function of the BM-Cyclin is to facilitate the accurate and reliable quantification of target analytes in a sample. The equipment utilizes advanced detection techniques to provide researchers and clinicians with essential data for their analyses.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Palbociclib, by Merck Group (1 mentions)
Dasatinib, by Selleck Chemicals (1 mentions)
Ribociclib, by Selleck Chemicals (1 mentions)
Canertinib, by Selleck Chemicals (1 mentions)
Abemaciclib, by MedChemExpress (1 mentions)
H1299, by Korean Cell Line Bank (1 mentions)
H1703, by Korean Cell Line Bank (1 mentions)
H1792, by Korean Cell Line Bank (1 mentions)
H1975, by Korean Cell Line Bank (1 mentions)
Hcc2108, by Korean Cell Line Bank (1 mentions)
Sk lu 1, by Korean Cell Line Bank (1 mentions)
Mycoplasma contamination, by Eurofins (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Rpmi glutamax, by Thermo Fisher Scientific (1 mentions)
Plasmocure, by InvivoGen (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Nutrient mixture f 12 dmem f 12, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
Cal27, by American Type Culture Collection (1 mentions)
16 protocols using bm cyclin
1
Cell Line Characterization and Inhibitor Screening
2
Cell Culture of Human Lung ADC
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Human lung ADC cell lines were obtained from the American Type Culture Collection (H1792) or the Korean Cell Line Bank (H23, H358, H1299, H1703, H1792, H1975, HCC1171, HCC2108, and SK-LU-1). All cells were grown at 37°C with 5% CO2 in RPMI-1640 or Dulbecco's modified Eagle's medium (HyClone) containing 10% fetal bovine serum (HyClone) and 1% penicillin/streptomycin (HyClone). All cells were monitored for Mycoplasma contamination using a PCR-based method for detection. Detected Mycoplasma were eliminated using BM cyclins (Roche).
3
Culturing and Embedding Human Breast Cancer Cells
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Human breast-cancer cell lines were obtained from the American Type Culture Collection and Korean Cell Line Bank. All cell lines were cultured according to the manufacturer's recommendations. All cells were screened for Mycoplasma contamination using a PCR-based detection method, and any identified contamination was eradicated using BM cyclins (Roche). Agarose pellets of human breast-cancer cell lines were embedded in a paraffin block.
4
Cell Culture Protocol for Various Cell Lines
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All cells were cultured in a 5% CO2 atmosphere at 37°C. MRC-5 (ECACC 05090501), HFF (HF-99/7 kindly provided by Dieter Neumann-Haefelin and Valeria Kapper-Falcone, Institute of Virology, Freiburg Germany), 293 T-CD20 (kindly provided by Irvin Chen, UCLA USA [Morizono et al., 2010 (link)]), BJ-Her2 (BJ-5ta foreskin fibroblasts [ATCC CRL-4001] stably expressing Her2/Erbb2 (NM_004448), lentiviral transduction as in Halenius et al., 2011 (link)), and Hela cells (ATCC CCL-2) were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% (vol/vol) fetal calf serum (FCS, Biochrom). BW5147 mouse thymoma cells (kindly provided by Ofer Mandelboim, Hadassah Hospital, Jerusalem, Israel) were maintained at 3 × 105 to 9 × 105 cells/ml in Roswell Park Memorial Institute medium (RPMI GlutaMAX, Gibco) supplemented with 10% (vol/vol) FCS, sodium pyruvate (1×, Gibco) and β-mercaptoethanol (0.1 mM, Sigma). Cells were used when tested negative for mycoplasma contamination (Eurofins). If positive, cells were treated using Plasmocure (Invivogen) or BM-Cyclin (Roche) as instructed by the supplier.
5
Establishment and Culture of OSCC Cell Lines
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The OSCC cell lines, SCC4, CAL27, and HSC3 were provided by Cara Gonzales at UTHSCSA, and were purchased from American Type Culture Collection (ATCC). The remaining OSCC cell lines, SCC9, SCC15, SCC25, SCC61, SCC131, SCC1352, and VU1729 were donated by Dr. Stephen Brant at Vanderbilt University. Cell lines were tested for mycoplasma and if positive, were treated with BM Cyclin (Roche). Cell lines were cultured in 50% Dulbecco's modification of Eagle's medium and 50% Nutrient Mixture F12 (DMEM/F12) (ThermoFisher Scientific), supplemented with 10% fetal bovine serum (FBS) (Hyclone Laboratories) and 1% penicillin/streptomycin (Mediatech). Cells were maintained at 37°C with 5% CO2.
6
Hematopoietic Cancer Cell Line Panel
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A panel of 21 hematopoietic cancer cell lines and primary cells from patients (listed in Table S1 and S2 ), was evaluated. Escherichia coli and Saccharomyces cerevisiae strains were kindly supplied by Dr Philippe Hauser (Institute of Microbiology, Lausanne University Hospital, Lausanne, Switzerland). All cells were cultured in RPMI (Invitrogen AG, 61870-01) supplemented with 10% heat inactivated fetal calf serum (Amimed, 2-01F30-I) and 1% penicillin/streptomycin at 37 °C (Amimed, 4-01F00-H) in a humidified atmosphere of 95% air and 5% CO2. To eliminate mycoplasma from cell culture, mycoplasma-infected leukemic cells, were cultured in the medium (as mentioned above), supplemented with BM-cyclin (Roche, Mannheim, Germany; Cat. No. 10799050001) according to the manufacturer’s instructions.
7
Intracellular Signaling in Engineered HEK293 Cells
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HEK293 cells (American Type Culture Collection) stably expressing FLAG-tagged β1AR or β2AR are maintained and transfected as previously described33 (link), 49 (link). Cells were periodically treated with BMCyclin (Roche) to avoid mycoplasma contamination. Cells were incubated overnight in serum-free medium supplemented with 0.1% BSA, 10 mM HEPES and 1% penicillin–streptomycin and pretreated with pertussis toxin (200 ng per ml, overnight), H89 (10 μM, 30 min) or propranolol (10 μM, 30 min) before ligand stimulation. HEK293 cells stably expressing β1AR-FRET sensor were used for the FRET experiments.
8
Itraconazole Modulates Hemangioma Endothelial Cells
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HemECs were obtained from proliferative IH tissues of 3 patients, the age of the 3 patients was less than 6 months. The patients underwent surgical operation at the Department of Oral and Maxillofacial Surgery of Qilu Hospital, Shandong University (Jinan, China). Human umbilical vein endothelial cells (HUVECs) were supplied by the American Type Culture Collection (Manassas, Virginia, USA). Cells at passages 3–7 were used in the following experiments. Cells used in studies were treated with the mycoplasma removal reagent BM-Cyclin (Roche, Germany) to ensure mycoplasma negativity before treatment. Dulbecco's modified Eagle's medium (Gibco, USA) containing 10% fetal bovine serum (HyClone, USA) and 1% penicillin/streptomycin (Gibco) was used to cultivate the cells. Itraconazole was obtained from Sigma-Aldrich. Itraconazole was dissolved in dimethyl sulfoxide (DMSO) to prepare the different concentrations of Itraconazole. The final concentration of DMSO in the medium was 0.1%. Cells treated with 0.1% DMSO were used as the control group. Recombinant human sonic hedgehog protein (rhSHH) was obtained from GenScript (Nanjing, China). All of the experiments were performed in triplicate wells and each experiment was performed in triplicate. Qilu Hospital, Shandong University, provided ethical permission for this investigation (Approval number: KYLL-202008-164).
9
Culturing and Treating Mouse Neuroblastoma Cells
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Hippocampus-derived mouse neuroblastoma HT22 cells (David Schubert, Salk Institute, La Jolla, CA) and HEK293T cells (ATCC, CRL-3,216) were cultured in Dulbecco’s modified Eagle’s medium (Gibco, 11,965-092) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, 12,306 C), 1% penicillin/streptomycin (P/S) (Gibco, 15,140–122), and BM-cyclin (Roche, 10799050001). Cells were kept at 37 °C with 5% CO2 levels. For treatment of H2O2, cells were supplemented with 1% FBS during the duration of H2O2 exposure.
10
Culturing Cell Lines for Experimental Assays
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Mouse hippocampus-derived neuroblastoma cells HT22, mouse embryonic fibroblast-derived NIH3T3, and tetracycline-inducible human embryonic kidney 293 cells overexpressing tau P301L (iHEK P301L) were cultured in Dulbecco’s modified Eagle’s medium (DMEM 1X) (Gibco, 11965-092) supplemented with 10% fetal bovine serum (FBS) (Sigma, 12306C) and 1% penicillin-streptomycin (P/S) (Gibco, 15140-122), and BM cyclin (Roche, 10799050001). iHEKP301L cells were induced with 1 μg/mL tetracycline when seeded, and cells were grown for 48 h before performing experimental assays. Cells were maintained at 37°C with 5% CO2 levels.
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