The human castration-resistant prostate cancer cell lines PC-3 and DU145 cells were preserved in RPMI1640 (Welgene, Daegu, Korea), supplemented with 10 % FBS and 1 % antibiotics (Welgene, Daegu, Korea). The human androgen responsive prostate cancer cell line LNCaP was maintained in RPMI1640, supplemented with 25 % HEPES (Welgene, Daegu, Korea), 10 % FBS and 1 % antibiotics (Welgene, Daegu, Korea). Normoxically conditioned cells were cultured in a 5 % CO2 incubator at 37 °C. The cells cultured under hypoxia were grown in a hypoxic chamber (Forma Scientific, Marietta, OH, USA) containing 1 % oxygen, 5 % carbon dioxide, and 94 % nitrogen at 37 °C.
Hepes
Manufactured by Welgene
Sourced in Switzerland
HEPES is a commonly used buffer solution used in cell culture and biochemical applications. It helps maintain a stable pH in the desired range, typically between 7.2 and 7.5, which is essential for the growth and survival of cells or the stability of biological molecules.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Welgene (2 mentions)
L glutamine, by Welgene (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dmem f12 media, by Welgene (1 mentions)
Insulin transferrin selenium solution, by Thermo Fisher Scientific (1 mentions)
Mle 12 cells, by American Type Culture Collection (1 mentions)
Penicillin streptomycin antibiotics, by Welgene (1 mentions)
Atplite system, by PerkinElmer (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Duchefa Biochemie (1 mentions)
Fetal bovine serum (fbs), by Welgene (1 mentions)
Ls202 02, by Welgene (1 mentions)
Streptomycin, by Welgene (1 mentions)
Fugene, by Promega (1 mentions)
Penicillin, by Welgene (1 mentions)
Human recombinant epidermal growth factor egf, by Thermo Fisher Scientific (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Welgene (1 mentions)
Fetal bovine serum (fbs), by GenDEPOT (1 mentions)
B16 cell line, by American Type Culture Collection (1 mentions)
7 protocols using hepes
1
Culturing Prostate Cancer Cell Lines
2
Cell Culture Protocols for RPE, Neurons, Macrophages, and Alveolar Epithelial Cells
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Human retinal pigment epithelial (RPE) cells were kindly provided by Dr. Jeong Hun Kim (College of Medicine, Seoul National University, Seoul, Korea). The murine hippocampal neuronal cell line HT-22 was kindly provided by Dr. Dong Gyu Jo (College of Pharmacy, Sungkyunkwan University, Suwon, Korea). The murine macrophage cell line RAW 264.7 was kindly provided by Dr. Sang Kook Lee (Seoul National University). Murine alveolar epithelial MLE-12 cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). RPE, HT-22, and RAW 264.7 cells were cultured in DMEM supplemented with 10% FBS and antibiotics (all from Welgene, Inc., Gyeongssan, Korea). MLE12 cells were cultured in HITES medium [DMEM-F12 media (Welgene) containing 1x insulin-transferrin-selenium solution (Thermo Fisher Scientific), 10 nM hydrocortisone, 10 nM β-estradiol, 10 mM HEPES, and 2 mM L-glutamine (Welgene)] supplemented with 2% FBS and antibiotics. Cells were incubated at 37°C with 5% CO2 in a humidified atmosphere.
3
Caco-2 Cell Viability Assay with AE-GBE
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Caco-2 cells were grown in 37 °C with 5% CO2 in MEM medium containing 25 mM HEPES (Welgene, Daegu, Korea), 10% FBS (Gibco, Paisley, UK), and 1% penicillin-streptomycin antibiotics (welgene). For cell toxicity assay on AE-GBE, the cells were treated with AE-GBE at different concentrations of 50, 100, 250, 500, and 1000 μg/mL for 24 h. Then, MTT (Duchefa Biochemie BV, Haarlem, Netherlands) of 0.5 mg/mL concentration was added to each plate and further cultured for 2 h; after removal of the supernatant, insoluble formazan crystals were dissolved in 2-propanol and measured at 540 nm (TECAN Group Ltd., Shanghai, China). As another method to analyse cell viability, ATP levels in the cells were measured with the Perkin-Elmer ATPLite system (PerkinElmer Life Sciences, Boston, MA, USA), according to the manufacturer’s instructions [20 (link)]. This system was based on the production of light caused by the reaction of ATP with added luciferase and D-luciferin. The attached mammalian cell lysis solution releases the adenine nucleotides and inactivates endogenous ATP degrading enzymes.
4
Gastric Cancer Cell Line Cultivation
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Human gastric cancer cell lines, MKN28 and AGS, were acquired from the Korean Cell Line Bank. The cells were used within ten passages. All cell lines were cultured in RPMI1640 (WelGENE, Gyeongsangbuk-do, Republic of Korea, LM011-07) with 25 mmol/L HEPES (WelGENE, BB001-01), 10% FBS (WelGENE), 100 U/mL penicillin (WelGENE, LS202-02), and 100 mg/mL streptomycin (WelGENE, LS202-02). Cells were incubated at 37 °C with 5% CO2. Transfection agent FuGENE (Promega, Wisconsin, USA, E5911) was used to facilitate transfection.
5
In Vitro Kidney Cell Culture
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For the in vitro investigation, the HK-2 Homo sapiens kidney tubular epithelial cell line and SV40 MES 13 mouse kidney glomerular mesangial cell line were purchased from American Type Culture Collection (Manassas, VA, USA). The HK-2 cells were cultured in complete growth media; keratinocyte serum-free medium supplemented with 0.05 mg/ml bovine pituitary extract, 5 ng/ml human recombinant epidermal growth factor (EGF) and 5 µg/ml gentamicin (Invitrogen Life Technologies, Carlsbad, CA, USA). The SV40 MES 13 cells were maintained in a 3:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium (Welgene, Daegu, Korea) with 14 mM HEPES (Welgene) supplemented with 5% fetal bovine serum (Invitrogen Life Technologies) and penicillin/streptomycin (100 U/ml; Invitrogen Life Technologies) at 37°C in 5% CO2 in a humidified incubator.
6
Culturing Melanoma and Melanocyte Cells
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The MNT-1 cell line (human melanoma) was kindly provided by Dr. Ai-Young Lee at Dongguk University, who originally received it as a gift from Dr. Vincent J. Hearing from the National Institutes of Health. The cells were cultured at 37 °C in EMEM (Gibco, Waltham, MA, USA) with 20% fetal bovine serum (FBS; GenDepot, Katy, TX, USA), 10% high-glucose DMEM (Lonza, Basel, Switzerland) as high energy source, 20 mM HEPES, and penicillin–streptomycin (Welgene, Gyeongsan, Republic of Korea). Normal human melanocytes were purchased from the Korean Cell Line Bank and were maintained at 37 °C in Medium 254 (Cascade Biologics, Waltham, MA, USA) with human melanocyte growth supplement (Life Technologies, Waltham, MA, USA) and penicillin–streptomycin. 2'-FL and DMP were obtained from Sigma-Aldrich (St. Louis, MO, USA). The B16 cell line (mouse melanoma) was purchased from the ATCC (Manassas, VA, USA) and was cultured in DMEM containing 10% FBS and 1% penicillin–streptomycin at 37 °C in a 5% CO2 atmosphere.
7
Investigating Vismodegib Effects on Intrahepatic CCC Cells
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Human intrahepatic CCC cell lines, SNU1079 and SNU-245 were purchased from the Korean Cell Line Bank. The cells were maintained at 37 °C in a humidified atmosphere of 5% CO2. Cells were cultured in RPMI1640 with L-glutamine (300 mg/L), 25 mM HEPES and 25 mM NaHCO3 (Welgene, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA). Cells were plated 105 cells per well on 6 well plate one day prior to drug treatment. Cells were treated with vismodegib at concentrations of 0, 1, 5, 10, and 20 μM. Cells were harvested at 0, 24, 48, and 72 h after the treatment and then used for cell viability assay (EZ-CYTOX; DoGenBio, Seoul, Korea).
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