Human VCaP (ATCC® CRL2876™), HCT-116 (ATCC® CCL247™), MIA-PaCa-2 (ATCC® CRL-1420™), HCC1954 (ATCC® CRL-2338™), and 786-O (ATCC® CRL-1932™) cells were a gift from Dr. Goutham Narla at the Case Comprehensive Cancer Center, Cleveland. VCaP and HCT-116 were gifted to Hera BioLabs in April 2017. MIA-PaCa-2, HCC1954, and 786-O were gifted to Hera BioLabs in June 2018. All cell lines were originally purchased from ATCC. VCaP and MIA-PaCa-2 cells were grown in Advanced DMEM (ThermoFisher #11995065) with 10% fetal bovine serum (Atlanta Biologicals # S12450) and 1% penicillin and streptomycin solutions (Cat# 15140–122, ThemoFisher). HCT-116 cells were grown in McCoy’s 5a Medium Modified (ATCC #30–2007) supplemented with 10% fetal bovine serum (Atlanta Biologicals # S12450) and 1% penicillin and streptomycin solutions (Cat# 15140–122, Themofisher). HCC1954 and 786-O cells were grown in RPMI 1640 (ThermoFisher # A1049101) with 10% fetal bovine serum (Atlanta Biologicals # S12450) and 1% penicillin and streptomycin solutions (Cat# 15140–122, ThemoFisher). All the cells were grown in a humidified incubator at 37°C with 5% CO2. All cells lines underwent monthly testing for mycoplasma contamination (Lonza, LT07-710) and STR testing at later passages.
S12450
Manufactured by Thermo Fisher Scientific
Sourced in United States
The S12450 is a high-precision analytical instrument designed for laboratory use. It provides accurate measurements and consistent performance for various applications. The core function of the S12450 is to enable precise analysis and data collection within a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Sv30010, by Thermo Fisher Scientific (2 mentions)
Advanced dmem, by Thermo Fisher Scientific (1 mentions)
Mia paca 2, by Thermo Fisher Scientific (1 mentions)
Lt07 710, by Lonza (1 mentions)
Mia paca 2, by American Type Culture Collection (1 mentions)
Hcc1954, by Thermo Fisher Scientific (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Hcc1954, by American Type Culture Collection (1 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rf 6a, by American Type Culture Collection (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
5 protocols using s12450
1
Cell Culture Protocols for Cancer Research
2
Cell Line Authentication and Culturing
3
Cell Line Authentication and Culturing
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All cell lines used in the study were obtained from American Type Culture Collection (ATCC), cultured under standard conditions and were confirmed to be free of mycoplasma. Parental cell lines and cell lines generated during the study were authenticated by ATCC Cell Line Authentication and were maintained in RPMI media (Fisher Scientific, 27-016-021) supplemented with 10% FBS (Atlanta Biologicals, S12450) and 1% penicillin/streptomycin (Fisher, SV300-10). Cell lines generated during this study, EKVX-pmiR and H322M-pmiR, were continuously cultured in media containing 16 or 8 μg/mL G418 (Fisher, 10-131-027), respectively. During the screen, media was changed to phenol red free RPMI media (Life Technologies, 11835030) supplemented with 10% FBS and 1% penicillin/streptomycin.
4
Culturing Intracellular Pathogens in Cell Lines
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The E. chaffeensis Arkansas strain1 (link) was cultured in THP-1 cells (ATCC, TIB-202),122 (link) and the A. phagocytophilum HZ strain123 (link) was cultured in HL-60 cells (ATCC, CCL-240) in RPMI 1640 medium (Mediatech, 10-040-CV) supplemented with 10% fetal bovine serum (Atlanta Biologicals, S12450) and 2 mM l -glutamine (Gibco, 25030). RF/6A cells (ATCC, CRL-1780) were cultured in advanced minimal essential medium (Gibco, 12492) supplemented with 10% fetal bovine serum and 2 mM l -glutamine. The method of synchronous culture of E. chaffeensis in HL-60 cells was similar to those described previously.9,43 (link) Host cell-free E. chaffeensis was used to initiate infection as previously described.9,43 (link) HEK293 cells (ATCC, CRL-1573) and canine histiocytic leukemia DH82 cells were cultured in DMEM (Dulbecco's minimal essential medium; Mediatech, 10-013-CV) supplemented with 10% fetal bovine serum and 2 mM l -glutamine.1 (link) Cultures were incubated at 37°C under 5% CO2 in a humidified atmosphere.
5
Cell Culture of HEK293T, Jurkat, and J-Lat
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HEK293T cells were maintained in DMEM (Corning 10017CV, Corning, NY, USA) containing 10% Fetal Bovine Serum (FBS; bio-techne S12450) and 100 U/mL Penicillin/Streptomycin (P/S; Gibco 15140163, Gaithersburg, MD, USA). Jurkat T cells and J-Lat 6.3 [43 (link)] cell lines were maintained in RPMI 1640 (Gibco 11875135, Gaithersburg, MD, USA) containing 10% heat-inactivated FBS, P/S. Cells were incubated at 37 °C, 5% CO2. All cell lines were obtained from ATCC (Manassas, VA, USA).
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