hA1 and hA3 AR functional experiments were carried out in CHO-A1 and CHO-A3#18 cell line, respectively. The day before the assay, the cells were seeded on the 96-well culture plate (Falcon 353072). The cells were washed with wash buffer (Mixture F12 Ham’s (Sigma N6658) for hA1 AR; Dulbecco’s modified eagle’s medium nutrient mixture F-12 ham (Sigma D8062) for hA3 AR; 25mM Hepes; pH = 7.4). Wash buffer was replaced by incubation buffer (Mixture F12 Ham’s (Sigma N6658) for hA1 AR; Dulbecco’s modified eagle’s medium nutrient mixture F-12 ham (Sigma D8062) for hA3 AR; 25 mM Hepes, 20 µM Rolipram (Sigma R6520); pH = 7.4). The cells were pre-incubated at 37 °C for 15 min. Then, test compounds and 5′-(N-Ethilcarboxamido) adenosine (NECA) as reference compound (Sigma E2387) were added and incubated at 37 °C for 10 min. FSK (Sigma F3917) was added and incubated at 37 °C for 5 min. After incubation, the amount of cAMP was determined using cAMP Biotrak Enzyme immunoassay (EIA) System Kit (GE Healthcare RPN225).
Ham s f 12
Manufactured by Merck Group
Sourced in United States, United Kingdom, Italy, Japan, Germany
Ham's F-12 is a cell culture medium designed for the cultivation of various cell types, including Chinese Hamster Ovary (CHO) cells, fibroblasts, and other mammalian cell lines. It provides the necessary nutrients and supplements to support the growth and maintenance of these cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (16 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (14 mentions)
Penicillin, by Merck Group (14 mentions)
Streptomycin, by Merck Group (13 mentions)
Hydrocortisone, by Merck Group (11 mentions)
L glutamine, by Merck Group (11 mentions)
Insulin, by Merck Group (10 mentions)
Penicillin streptomycin, by Merck Group (9 mentions)
Fetal calf serum (fcs), by Euroclone (8 mentions)
Rpmi 1640 medium, by Euroclone (8 mentions)
Streptomycin, by Thermo Fisher Scientific (8 mentions)
Fetal bovine serum (fbs), by Merck Group (8 mentions)
Bxpc 3, by American Type Culture Collection (7 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (7 mentions)
Cholera toxin, by Merck Group (7 mentions)
Glutamax, by Thermo Fisher Scientific (7 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
Glutamine, by Merck Group (6 mentions)
Mcf 7, by American Type Culture Collection (5 mentions)
Dexamethasone (dex), by Merck Group (5 mentions)
110 protocols using ham s f 12
1
Adenosine Receptor Functional Assay in CHO Cells
2
Comparative Study of Human Uterine Cell Lines
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SKN, which is a human uLMS cell line, and MES-SA, which is human uterine sarcoma cell line, were used in this study. SKN was purchased from Health Science Research Resources Bank (HSRRB, Osaka, Japan), and MES-SA was purchased from American type culture collection (ATCC, Virginia, USA). SKN cells were cultured in Ham's F 12 (Sigma-Aldrich Japan K.K., Tokyo, Japan), and MES-SA cells were cultured in McCoy's 5a medium (ATCC). Both media were supplemented with 10% heat-incubated fetal bovine serum (FBS). The cells were seeded at a density of 5×104 cells/well in a six-well plate, and incubated at 37°C in a humidified 5% CO2 incubator for 5 days. The cells were trypsinized and counted by a cell counter (Vi-CELL XR; Beckman Coulter, Tokyo, Japan) at each time point, as reported previously (36 (link)).
For TGF-β treatment, cells were cultured in Ham's F12 and McCoy's 5a medium supplemented with 10% FBS containing 100 or 500 pg/ml TGF-β1 (Sigma-Aldrich Japan K.K.) for 24 h. In order to block TGF-β signaling, SB431542 (WAKO, Tokyo, Japan), which is a TGF-β type I receptor-selective blocker, was dissolved at a concentration of 10 µM in dimethylsulfoxide (DMSO). Cells were seeded at a density of 5×104 cells/well in a six-well plate and cultured for 24 h and then cultured with new medium supplemented with 10 µM SB431542 for more 48 h.
For TGF-β treatment, cells were cultured in Ham's F12 and McCoy's 5a medium supplemented with 10% FBS containing 100 or 500 pg/ml TGF-β1 (Sigma-Aldrich Japan K.K.) for 24 h. In order to block TGF-β signaling, SB431542 (WAKO, Tokyo, Japan), which is a TGF-β type I receptor-selective blocker, was dissolved at a concentration of 10 µM in dimethylsulfoxide (DMSO). Cells were seeded at a density of 5×104 cells/well in a six-well plate and cultured for 24 h and then cultured with new medium supplemented with 10 µM SB431542 for more 48 h.
3
Diverse Carcinoma Cell Lines Maintenance
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Human lung (A549), breast (MCF-7), colon (HCT-15), and pancreatic (BxPC3) carcinoma cell lines along with melanoma (A375) cells were obtained by American Type Culture Collection (ATCC, Rockville, MD, USA). A431 are human cervical carcinoma cells kindly provided by Professor F. Zunino (Division of Experimental Oncology B, Istituto Nazionale dei Tumori, Milan, Italy). The 2008 cells and cisplatin-resistant variant, C13*, are human ovarian adenocarcinoma cell lines that were kindly provided by Professor G. Marverti (Department of Biomedical Science of Modena University, Italy). Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (EuroClone, Milan, Italy), antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin) and 2 mM l-glutamine: (i) RPMI-1640 medium (EuroClone) for MCF-7, A431, BxPC3, 2008 and C13* cells; (ii) F-12 HAM’S (Sigma Chemical Co.) for A549 cells; (iii) DMEM for A375 cells.
4
Diverse Cancer Cell Lines Cultivation
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Human breast (MCF-7), colon (HCT-15), lung (A549) and pancreatic (BxPC3) carcinoma cell lines together with melanoma (A375) cells were obtained from American Type Culture Collection (ATCC, Rockville, MD). The human ovarian cancer cell line 2008 and its cisplatin resistant variant, C13*, were kindly provided by Prof. G. Marverti (Department of Biomedical Science of Modena University, Italy). Human cervical carcinoma (A431) cells were kindly provided by Prof. F. Zunino (Division of Experimental Oncology B, Istituto Nazionale dei Tumori, Milan, Italy). Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (Euroclone, Milan, Italy), antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin) and 2 mM L-glutamine: (i) RPMI-1640 medium (Euroclone) for 2008, C13*, MCF-7, HCT-15, A431 and BxPC3 cells, (ii) F-12 HAM’S (Sigma Chemical Co.) for A549 cells, iii) DMEM (Sigma Chemical Co.) for A375 cells.
5
Establishment and Culture of Carcinoma Cell Lines
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Human lung (A549), breast (MCF-7), pancreas (BxPC3), and colon (LoVo) carcinoma cell lines along with melanoma (A375) were obtained from American Type Culture Collection (ATCC, Rockville, MD). Human non-tumor embryonic kidney HEK293 cells were obtained from European Collection of Cell Cultures (ECACC, Salisbury, UK). Human cervical carcinoma A431cells were kindly provided by Prof. F. Zunino (Division of Experimental Oncology B, Istituto Nazionale dei Tumori, Milan, Italy). Human ovarian cancer 2008 cells were kindly provided by Prof. G. Marverti (Dept. of Biomedical Science of Modena University, Italy). Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (Euroclone, Milan, Italy), antibiotics (50 units ∙ mL−1 penicillin and 50 μg ∙ mL−1 streptomycin) and 2 mM l-glutamine: i) RPMI-1640 medium (Euroclone) for MCF-7, A431, BxPC3 and 2008 cells; ii) F-12 HAM’S (Sigma Chemical Co.) for A549 and LoVo cells; iii) D-MEM medium (Euroclone) for HEK293 cells.
6
Cell Lines for Oncology Research
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Human lung (A549), colon (LoVo and HCT-15) and pancreatic (BxPC3, PSN-1) carcinoma cell lines, along with melanoma (A375) cells, were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). Embryonic kidney HEK293 cells were obtained from the European Collection of Cell Cultures (ECACC, Salisbury, UK). A431 are human cervical carcinoma cells kindly provided by Prof. F. Zunino (Division of Experimental Oncology B, Istituto Nazionale dei Tumori, Milan, Italy). The 2008 cells and cisplatin-resistant variant, C13*, are human ovarian adenocarcinoma cell lines that were kindly provided by Prof. G. Marverti (Dept. of Biomedical Science of Modena University, Italy). The LoVo OXP cells were derived, using a standard protocol, by growing LoVo cells in increasing concentrations of oxaliplatin and following nine months of selection of resistant clones. Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (Euroclone, Milan, Italy), antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin) and 2 mM l-glutamine: i) RPMI-1640 medium (Euroclone, Milan, Italy) for BxPC3, PSN-1, 2008 and C13* cells; ii) F-12 HAM′S (Sigma Chemical Co., Darmstadt, Germany) for A549, LoVo and LoVo OXP cells; and iii) DMEM for A375 and HEK293 cells.
7
CHO-K1 Cell Culture Protocol
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Chinese hamster ovary (CHO-K1) cells purchased from ECACC (catalogues number 85051005, lot 12G006, passage number P+7) were cultured (37°C, 5% CO 2 ) in F-12 Ham's (Sigma Aldrich) supplemented with 10% fetal calf serum (v/v), 100 U/mL penicillin, 100 µg/mL streptomycin, 0.3 mg/mL L-glutamine (Gibco). Fetal calf serum was used to adhere to the original clustering protocol (Isbrucker et al., 2016) , though replacement of this medium component is envisaged in the future. Cells were passaged every 2-4 days using trypsin-EDTA, and cells of passage numbers 12-20 were used for the experiments.
8
Establishing Carcinoma Cell Line Cultures
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Human breast (MCF-7), colon (HCT-15), and pancreatic (BxPC3, MIA Paca-2, AsPC-1, Capan-1, PANC-1) carcinoma cell lines along with melanoma (A375) cells and nontransformed embryonic kidney (HEK293) were obtained from American Type Culture Collection (ATCC, Rockville, MD). The human ovarian 2008 cancer cell line was kindly provided by Prof. G. Marverti (Dept. of Biomedical Science of Modena University, Italy). Human cervical carcinoma (A431) cells were kindly provided by Prof. F. Zunino (Division of Experimental Oncology B, Istituto Nazionale dei Tumori, Milan, Italy). Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (Euroclone, Milan, Italy), antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin) and 2 mM L-glutamine: (i) RPMI-1640 medium (Euroclone) for 2008, MCF-7, HCT-15, A431, AsPC-1 and BxPC3 cells; (ii) F-12 HAM'S (Sigma Chemical Co.) for A549 cells; iii) DMEM (Sigma Chemical Co.) for MIA Paca-2, PANC-1, A375 and HEK293 cells; IMDM (Sigma Chemical Co.) for Capan-1 cells.
9
Maintenance of Carcinoma and Melanoma Cell Lines
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The human pancreatic (BxPC3) and colon (LoVo) carcinoma and melanoma (A375) cell lines were obtained from American Type Culture Collection (ATCC, Rockville, MD). The human squamous cervical cancer cell line (A431) was kindly provided by Prof. G. Zunino (Division of Experimental Oncology B, Istituto Nazionale dei Tumori, Milan, Italy).
Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (Euroclone, Milan, Italy), antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin), and 2 mM L-glu- tamine: (i) RPMI-1640 medium (Euroclone) for BxPC3 and A431 cells; (ii) DMEM for A375 cells; (iii) F-12 HAM'S (Sigma Chemical Co.) for LoVo cells.
Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (Euroclone, Milan, Italy), antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin), and 2 mM L-glu- tamine: (i) RPMI-1640 medium (Euroclone) for BxPC3 and A431 cells; (ii) DMEM for A375 cells; (iii) F-12 HAM'S (Sigma Chemical Co.) for LoVo cells.
10
Culturing Various Cancer Cell Lines
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Human breast (MCF-7), colon (HCT-15 and LoVo), lung (A549), and pancreatic (BxPC3) carcinoma cell lines along with melanoma (A375) cells were obtained from American Type Culture Collection (ATCC, Rockville, MD). The human ovarian cancer cell line 2008 and its cisplatin resistant variant, C13*, were kindly provided by Prof. G. Marverti (Dept. of Biomedical Science of Modena University, Italy). The human colon carcinoma LoVo-OXP cells were derived, using a standard protocol, by growing LoVo cells in increasing concentrations of oxaliplatin and following 17 months of selection of resistant clones, as previously described. [28] Cell lines were maintained in the logarithmic phase at 37 °C in a 5% carbon dioxide atmosphere using the following culture media containing 10% fetal calf serum (Euroclone, Milan, Italy), antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin) and 2 mM L-glutamine: (i) RPMI-1640 medium (Euroclone) for 2008, C13*, MCF-7, HCT-15 and BxPC3 cells; (ii) F-12 HAM'S (Sigma Chemical Co.) for A549, LoVo and LoVo-OXP cells; iii) DMEM (Sigma Chemical Co.) for A375 cells.
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