The human colon cancer cell lines HT-29, Caco-2, RKO, HCT116 and Colo205 were purchased from cell bank (Shanghai Institute of Biological Sciences, Shanghai, China). Colo205, HT-29 and RKO grew as monolayers in Dulbecco’s modified Eagles medium (DMEM) containing 10% fetal calf serum (Gibco, Grand Island, NY, USA) in a 5% CO2, 95% at 37 °C. Caco-2 grew in DMEM containing 20% fetal calf serum (Gibco). HCT116 grew in McCoy’5 A (Sigma, Saint Louis, MO, USA) containing 10% fetal calf serum (Gibco). Oxaliplatin was purchased from Meilun Biological Co., Ltd. (Dalian, China). Oxaliplatin was dissolved at 2000 μM in 5% GS, as stocks and stored at −20 °C. Oxaliplatin stocks were diluted at a series of concentrations in serum-free DMEM immediately prior to use in the in vitro experiments. For the in vivo studies, oxaliplatin was dissolved at 0.625 g/l in 5% GS and 5% GS was used as the solvent control. N-Acetyl-cysteine (NAC) was purchased from Sigma. NAC was dissolved at 100 mM in serum-free DMEM, as stocks and stored at −20 °C.
Mccoy 5a
Manufactured by Merck Group
Sourced in United States
The McCoy 5A is a versatile laboratory equipment designed for various applications. It functions as a precise and reliable instrument for measurement, analysis, and data collection tasks within a controlled laboratory environment. The core purpose of the McCoy 5A is to provide accurate and consistent results, enabling researchers and scientists to conduct their experiments and investigations effectively.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Mia paca 2, by American Type Culture Collection (2 mentions)
H45 hypoxystation, by Don Whitley Scientific (2 mentions)
Bxpc 3, by American Type Culture Collection (2 mentions)
Capan 2, by American Type Culture Collection (2 mentions)
Rpmi 1640, by Merck Group (2 mentions)
Dulbecco s modified eagle s medium, by Merck Group (2 mentions)
Nu7441, by MedChemExpress (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Panc 1, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Colo205, by Thermo Fisher Scientific (1 mentions)
N acetylcysteine nac, by Merck Group (1 mentions)
Caco 2, by Thermo Fisher Scientific (1 mentions)
Hct116, by Merck Group (1 mentions)
Ht 29, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Nissui Pharmaceutical (1 mentions)
Rpmi 1640 medium, by Nissui Pharmaceutical (1 mentions)
10 protocols using mccoy 5a
1
Colon Cancer Cell Lines Cultivation
2
Culturing Human Ovarian Cancer Cells
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Human ovarian cancer OVISE and JHOC5 cells were cultured in DMEM and RPMI‐1640 medium (Nissui) supplemented with 10% FBS, respectively. TOV21G cells were cultured in MCDB 105 (50%)/Medium 109 (Sigma) supplemented with 10% FBS. ES2, HCT116 (p53+/+), and HCT116 (p53−/−) cells were grown in McCoy 5A (Sigma) supplemented with 10% FBS.
3
Chemosensitivity Assay Protocol
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Media (DMEM, McCoy5a, RPMI), penicillin, streptomycin, FBS, L-glutamine, G418, along with GEM, doxorubicin, 5-fluorouracil, paclitaxel, procainamide, and ampicillin were purchased from Sigma-Aldrich (St. Louis, MO) and reconstituted as per manufacturer's instructions. Selumetinib (AZD6244) was purchased from ChemieTek (Indianapolis, IN) and reconstituted in 1X PBS at 10 mg/mL concentration, which formed a white, turbid suspension. GEM was dissolved in 1X PBS at 5 mg/mL. U0126 was purchased from Tocris Biosciences (Bristol, UK) and reconstituted in DMSO at 10 mM. 13C[15N2]-GEM (99% purity) was obtained from Moravek Biochemicals (Brea, CA). Methanol, acetonitrile (HPLC grade), formic acid, and acetic acid were purchased from Mallinckrodt Baker (Phillipsburg, NJ). Antibodies against P-gp, ENT1, MRP1, MRP5, β-actin, and secondary antibodies were purchased from Abcam (Cambridge, MA), and antibodies against phospho-ERK and total-ERK were purchased from Cell Signaling (Danvers, MA) and ARF6 from Abgent (San Diego, CA). The antibodies were previously published [35 (link)].
4
Culturing Pancreatic Cancer Cell Lines
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PANC-1 [American Type Culture Collection (ATCC), CRL-1469], BxPC-3 (ATCC, CRL-1687), SK-PC-3, and GP-9A cells were cultured in RPMI 1640 (Sigma-Aldrich). Capan-2 (ATCC, HTB-80) cells were cultured in McCoy 5A (Sigma-Aldrich). MIA PaCa-2 (ATCC, CRL-11268), GP-2A, GP-3A, GP-5A, GP-9A, GP-10A, and GP-16A cells were cultured in Dulbecco’s modified Eagle’s medium (Sigma-Aldrich). All media were supplemented with 2 to 10% fetal bovine serum (Gibco) and cultured under conditions of 5% CO2 in air at 37°C. For primary pancreatic cancer cells, 1× penicillin/streptomycin was added in the growth medium. For hypoxia, cells were transferred to H45 Hypoxystation (Don Whitley Scientific). The DNA-PK inhibitor NU7441 (10 μM, Medchem Express) was added at the time of seeding and replenished 1 hour before collecting the cells.
5
Culturing Cervical Cancer Cell Lines
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Cervical cancer cells HeLa, SiHa and ME 180 were procured from National Centre for Cell Science (Pune, India) and maintained in 5% CO2 atmosphere, at 37 °C and 95% humidity in DMEM (Gibco-BRL, Rockville, MD, USA; HeLa and SiHa) and McCoy-5A (Sigma Aldrich, St. Louis, MO, USA; ME 180) supplemented with 10% heat inactivated FBS (Gibco-BRL)
6
Cell Culture and Transfection Protocol
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Cells were cultured at 37°C in the presence of 5% CO2 in McCOY’5A or DMEM media (Sigma) for colon cancer and breast cancer cells, respectively. HCT116 (RRID:CVCL-0291 ), MDA-MB231 (RRID:CVCL-0062 ), MCF7 (RRID:CVCL-0031 ), LoVo (RRID:CVCL-0399 ), SW480 (RRID:CVCL-0546 ), SW620 (RRID:CVCL-0547 ), Colo205 (RRID:CVCL-0218 ) were purchased from ATCC. MDA-MB231 D3H2LN (RRID:CVCL-D257 ) were purchased from PerkinElmer. All cell lines were tested for mycoplasma contamination after thawing using Lonza mycoalert assay (Reference number: LT27-236). Transfections of p53 isoforms and siRNA were carried out using JetPEI kit (Qbiogen) and Interferin (Polyplus), respectively, according to the manufacturers’ instructions. For adhesion assays, non-adherent and adherent cells were collected 24 hr after transfection and counted using the Countess cell counting system (Invitrogen).
7
Optimizing Hematopoietic Cell Culture Conditions
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McCoy’5A (M-4892, Sigma), RPMI 1640 (31800-022, Gibco), Waymouth’s medium (31220023, Gibco), Iscove’s modified Dulbecco’s medium (IMDM) (12440-053, Gibco), orotic acid (02750-10G, Sigma), l -ascobic acid (A4544-100G, Sigma), Inosine 5′-monophosphate disodium salt (I4625-25G), l -glutathione reduced (G6013-100G, Sigma), HT supplement (11067-030, Gibco), cholesterol lipid concentrate (12531-016, Gibco), ACK red blood cell lysis buffer (A10492-01, Gibco), Pen/Strep solution (15140-122, Gibco), erythropoietin (EPO) (Exprex4000, Janssen-Cilag, Australia), Stem cell factor (SCF) (produced in-house at WEHI), Histopaque (10771, Sigma), Nycoprep Universal 60% solution (AN1106865, Axis-Shield).
8
Molecular Mechanisms of Cancer Progression
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McCoy 5A was purchased from Sigma. T4 DNA ligase was purchased from Fermentas. The pcDNA™6.2-GM/EmGFP-miR, pcDNA™6.2-GM/EmGFP-miR-neg-control plasmid and Lipofectamine 2000 were purchased from Invitrogen. Trizol reagent was purchased from Takara. RevertAid First Strand cDNA Synthesis kits were purchased from Thermo. Antibodies against VEGF, PTEN, AKT, and phospho-AKT were purchased from Abcam. Antibodies against GAPDH, ERK1/2 and phospho-ERK1/2 were purchased from Cell Signaling Technology. Cell counting kit-8 reagent was purchased from Boster. Transwell chambers were purchased from Costar. SYBR® Premix Ex Taq™ II was purchased from Takara. C1000™Thermal Cycler and S1000™Thermal Cycler were purchased from BIO-RAD. Flow cytometry from Beckman Coulter.
9
Pancreatic Cancer Cell Culture Conditions
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PANC-1 (ATCC, CRL-1469), BxPC-3 (ATCC, CRL-1687), SK-PC-3 and GP-9A cells were cultured in RPMI-1640 (Sigma). Capan-2 (ATCC, HTB-80) cells were cultured in McCoy 5A (Sigma). MIA PaCa-2 (ATCC, CRL-11268), GP-2A, GP-3A, GP-5A, GP-9A, GP-10A and GP-16A cells were cultured in DMEM (Sigma). All media was supplemented with 2%-10% fetal bovine serum (Gibco) and cultured under conditions of 5% CO2 in air at 37⁰C. For primary pancreatic cancer cells 1X penicillin/streptomycin was added in the growth media. For hypoxia, cells were transferred to a H45 Hypoxystation (Don Whitley Scientific). DNA-PK inhibitor Nu7441 (10 µM, Medchem Express) was added at the time of seeding and replenished one hour before collecting the cells.
10
Melanoma Cell Line Maintenance and Stimulation
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B16 (mouse melanoma), 13 MMAc (human melanoma; RIKEN, Tsukuba, Japan), and A2058 (human melanoma; JCRB, Osaka, Japan) cell lines were maintained in Dulbecco's modified Eagle's medium (Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS; Sigma) and 1% penicillin/streptomycin (100 U/mL; Life Technologies, Tokyo, Japan). The B16/BL6 cell line (mouse melanoma) 14 was maintained in RPMI 1640 medium (Sigma) with 10% FBS and 1% penicillin/streptomycin. The G-361 cell line (human melanoma; RIKEN) was maintained in McCoy5A (Sigma) with 10% FBS and 1% penicillin/streptomycin. Finally, the MeWo cell line (human melanoma; JCRB) was maintained in minimal essential medium (Sigma) with 10% FBS, 1% nonessential amino acids (Gibco, Grand Island, NY), and 1% penicillin/streptomycin.
Recombinant Gal-3 and TGF-b1 (R&D Systems, Minneapolis, MN), TGF-b receptor 1 inhibitor (LY364947; Merck Millipore, Darmstadt, Germany), and fibroblast growth factor 2 and epidermal growth factor (EGF) (Kurabo, Osaka, Japan) were used to stimulate the cells. For this, cells were starved in FBS-free medium for 4 to 8 hours before adding reagents in 24-well plates. Mouse Gal-3 siRNA and negative control siRNA were purchased from Sigma.
Recombinant Gal-3 and TGF-b1 (R&D Systems, Minneapolis, MN), TGF-b receptor 1 inhibitor (LY364947; Merck Millipore, Darmstadt, Germany), and fibroblast growth factor 2 and epidermal growth factor (EGF) (Kurabo, Osaka, Japan) were used to stimulate the cells. For this, cells were starved in FBS-free medium for 4 to 8 hours before adding reagents in 24-well plates. Mouse Gal-3 siRNA and negative control siRNA were purchased from Sigma.
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