Hek293ft

Manufactured by Merck Group

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The HEK293FT cell line is a human embryonic kidney cell line that has been engineered to express the SV40 large T antigen. This cell line is commonly used in various research applications, including virus production, protein expression, and gene transfection studies.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Hek293ft, by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Thp 1, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions) Geneticin, by Merck Group (1 mentions) Raw264, by American Type Culture Collection (1 mentions) Hek293t, by Merck Group (1 mentions) Venorgem mycoplasma pcr detection kit, by Minerva Biolabs (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Glutamine, by Euroclone (1 mentions) Hek293ft, by Euroclone (1 mentions) Penicillin streptomycin, by Euroclone (1 mentions) Dulbecco modified eagle medium (dmem), by Euroclone (1 mentions) Fetal bovine serum (fbs), by Euroclone (1 mentions) Rpmi 1640 medium, by Euroclone (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Screenfect a, by Fujifilm (1 mentions) Lenti x concentrator, by Takara Bio (1 mentions)

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HEK293FT cells (2 citations)

8 protocols using hek293ft

Culturing HEK293FT, RAW264.7, and THP-1 Cell Lines

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The human embryonic kidney cell line HEK-293FT (Thermo Scientific-Life Technologies, Waltham, MA, USA) was cultured in DMEM (Thermo Scientific-Gibco, Grand Island, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (GIBCO) with 100 U/mL penicillin and 100 µ/mL streptomycin at 37 °C in 5% CO₂. The HEK293FT cells were maintained with 500 μg/mL of geneticin (SIGMA ALDRICH, St. Louis, MO, USA). The murine macrophage cell line RAW264.7 (ATCC:TIB-71, American Type Culture Collection, Manassas, VA, USA) was cultured in DMEM (GIBCO) supplemented with 10% heat-inactivated fetal bovine serum (GIBCO) with 100 U/mL penicillin and 100 µ/mL streptomycin at 37 °C in 5% CO₂. The human monocytic leukemia cell line THP-1 (ATCC:TIB202TM) was cultured in RPMI (GIBCO) supplemented with 20% heat-inactivated fetal bovine serum (GIBCO) with 100 U/mL penicillin and 100 mg/mL streptomycin at 37 °C in 5% CO₂.
Murine primary macrophages were thioglycollate-elicited and removed from wild-type (WT) or TLR4-knockout (KO) C57BL/6 mice by peritoneal washing. Primary macrophages were dispensed into 6-well plates at a 2 × 10⁶ concentration for 1 hour to allow cells to adhere. Adherent cells were cultured for an additional 24 hours in DMEM.
(GIBCO) supplemented with 10% heat-inactivated fetal bovine serum (GIBCO) with 100 U/mL of penicillin and 100 µ/mL streptomycin at 37 °C in 5% CO₂.

Dias-Teixeira K.L., Calegari-Silva T.C., Medina J.M., Vivarini Á.C., Cavalcanti Á., Teteo N., Santana A.K., Real F., Gomes C.M., Pereira R.M., Fasel N., Silva J.S., Aktas B.H, & Lopes U.G. (2017). Emerging Role for the PERK/eIF2α/ATF4 in Human Cutaneous Leishmaniasis. Scientific Reports, 7, 17074.

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HEK293T and HEK293FT Cell Cultivation

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HEK293T (RRID:CVCL_0063) and HEK293FT (RRID:CVCL_6911) cell lines were obtained from Sigma-Aldrich (Saint Louis, MO) and Invitrogen (Carlsbad, CA)/ThermoFisher Scientific (Waltham, MA), respectively. The identity of cell lines was authenticated by internal SNP genotype profiling. The absence of mycoplasma contamination was regularly verified (Venor GeM Mycoplasma PCR Detection kit, Minerva Biolabs, Germany). Both cell lines were maintained in DMEM supplemented with 10% (v/v) fetal calf serum (AMIMED, United Kingdom), 2 mM L-glutamine, 1 mM sodium pyruvate and 0.1 mM MEM non-essential amino acids. Transient transfections were performed with a DNA mix containing the plasmids of interest using Lipofectamine 2000 (Invitrogen, Carlsbad, CA), according to the manufacturer's protocol.

Mesrouze Y., Bokhovchuk F., Meyerhofer M., Fontana P., Zimmermann C., Martin T., Delaunay C., Erdmann D., Schmelzle T, & Chène P. (2017). Dissection of the interaction between the intrinsically disordered YAP protein and the transcription factor TEAD. eLife, 6, e25068.

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Cell Culture Protocols for Cell Line Research

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U937, K562 and HL-60 cells (DSMZ) were cultured in RPMI 1640 medium (Euroclone, Italy) supplemented with 10% heat-inactivated FBS (Sigma-Aldrich, Italy), 1% glutamine (Euroclone) and 1% penicillin/streptomycin (Euroclone) at 37 °C and 5% CO2. HEK293-FT (Sigma-Aldrich) and HeLa cells (provided by M. Vermeulen) were grown in DMEM (Euroclone) supplemented with 10% FBS, 100 U/mL penicillin/streptomycin (Euroclone) and 6 mM (HEK293-FT) or 2 mM (HeLa) glutamine (Euroclone). They have been tested for mycoplasma contamination.

Di Costanzo A., Del Gaudio N., Conte L., Dell’Aversana C., Vermeulen M., de Thé H., Migliaccio A., Nebbioso A, & Altucci L. (2018). The HDAC inhibitor SAHA regulates CBX2 stability via a SUMO-triggered ubiquitin-mediated pathway in leukemia. Oncogene, 37(19), 2559-2572.

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Lentiviral Transduction and siRNA Knockdown in U87MG Cells

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Plasmids for CD133 were gifted by Dr. Young-Gyu Ko (Korea University). Third-generation lentiviral packaging vectors were transfected with PolyExpress™ (Excellgen Inc., USA) in HEK-293FT (Invitrogen, USA). Lentiviruses were concentrated with Lenti-X™ Concentrator (Clontech, Japan). U87MG cells were infected with the lentivirus produced from HEK-293FT.
To perform siRNA-mediated knockdown of IL-1β gene, two siRNAs (SASI_Hs01_00028205 and SASI_Hs02_00302835; Sigma Aldrich) were transfected in U87MG cells by using ScreenFect™-A (Wako Pure Chemical Industries, Japan), according to the manufacturer’s instructions. Cells were harvested 48 h after transfection.

Lee S.Y., Kim J.K., Jeon H.Y., Ham S.W, & Kim H. (2017). CD133 Regulates IL-1β Signaling and Neutrophil Recruitment in Glioblastoma. Molecules and Cells, 40(7), 515-522.

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Cell Culture Conditions for Burkitt's Lymphoma and Fibroblasts

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Ramos human Burkitt’s lymphoma B cells (CRL-1596, American Type Culture Collection, Manassas, VA, USA) were grown in RPMI 1640 medium supplemented with 2 mM L-glutamine and 10% heat-inactivated fetal bovine serum (HyClone, Logan, UT, USA). BALB/3T3 fibroblasts and HEK293 cells were obtained from ATCC (Manassas, VA, USA). HEK293FT human embryonic kidney cells (Thermo Fisher Scientific, San Jose, CA, USA). BALB/3T3 fibroblasts, HEK293 and HEK293FT were maintained in Dulbecco’s modified Eagle’s medium (Sigma, Saint Louis, MO, USA) supplemented with 10% fetal calf serum, 100 μg/mL streptomycin and 100 U/ml penicillin grown in a humidified incubator at 37 °C in 5% CO₂ and 95% air. 3T3/TetOn-AID cells^{10 (link)} were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% dialyzed fetal calf serum, 100 μg/mL streptomycin and 100 U/ml penicillin grown in a humidified incubator at 37 °C in 5% CO₂ and 95% air.

Al-Qaisi T.S., Su Y.C, & Roffler S.R. (2018). Transient AID expression for in situ mutagenesis with improved cellular fitness. Scientific Reports, 8, 9413.

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Culturing NSCLC and HEK-293FT Cell Lines

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The human NSCLC cell lines (PC9, SPC-A1, A549, H1299, H1975 and GLC-82) and human embryonic kidney cells (HEK-293FT) were all purchased from the ATCC. PC9, A549, H1299 and H1975 cells were grown in RPMI 1640 medium. SPC-A1, GLC-82 and HEK-293FT cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) medium supplemented with 10% fetal bovine serum and 100 IU/ml penicillin/streptomycin (Sigma). All cells were grown at 37 °C in a humidified 5% CO₂ atmosphere.

Xu M., Wang F., Li G., Wang X., Fang X., Jin H., Chen Z., Zhang J, & Fu L. (2019). MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC. Molecular Cancer, 18, 93.

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Cell Line Acquisition and Culture

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Human embryonic lung MRC5 fibroblasts and HeLa cells were obtained from European Collection of Authenticated Cell Cultures (ECACC 05011802 and 93021013) and HEK 293FT were purchased from Life Technologies (R700-07). Sqstm1 knockout (sqstm1^−/−) and wild type (Sqstm1^+/+) mouse embryonic fibroblasts (MEFs) were kindly provided by Dr. Eiji Warabi of the University of Tsukuba.¹⁵ (link)
Atg5-deficient (atg5^−/−) and wild-type (Atg5^+/+) MEFs and M5-7 MEFs were kindly provided by Dr Noboru Mizushima (University of Tokyo).^28,29 (link) All cells were grown in Dulbecco's modified Eagle's medium (Sigma, D6546) supplemented with 10% heat-inactivated fetal bovine serum (Biosera, FB-1001H), 5% penicillin/streptomycin (Invitrogen, 15140122) and 2 mM L-glutamine (Sigma, 59202C) in a humidified atmosphere containing 5% CO₂ at 37°C. Autophagy was abolished in M5-7 MEFs by treating with 1 μg/ml tetracycline for at least 4 d. HEK 293FT cells were maintained in 500 µg/ml G418 (Sigma, A1720) prior to transfection.

Hewitt G., Carroll B., Sarallah R., Correia-Melo C., Ogrodnik M., Nelson G., Otten E.G., Manni D., Antrobus R., Morgan B.A., von Zglinicki T., Jurk D., Seluanov A., Gorbunova V., Johansen T., Passos J.F, & Korolchuk V.I. (2016). SQSTM1/p62 mediates crosstalk between autophagy and the UPS in DNA repair. Autophagy, 12(10), 1917-1930.

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Culturing CCD841 and HEK293FT Cell Lines

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Normal human colonic epithelial cell line, CCD841, was purchased from American Type Culture Collection (ATCC, Manassas, VA, U.S.A.). The cell line was cultured in RPMI 1640 (Corning, Corning, NY, U.S.A.) supplemented with 10% FBS (Fisher Scientific, Pittsburgh, PA, U.S.A.) and 1% penicillin/streptomycin (Corning, Corning, NY, U.S.A.). Viral packaging cell line, HEK293FT, was purchased from ATTC (Manassas, VA, U.S.A.) and maintained in DMEM 4.5 g/L glucose, l-glutamine, sodium pyruvate (Corning, Corning, NY, U.S.A.) supplemented with 10% FBS and 1% penicillin/streptomycin. HEK293FT cells were used for transient transfection experiments where they were cultured for three passages in the presence and absence of excess iron in the form of ferric ammonium citrate (FAC) (Sigma, St. Louis, MO, U.S.A.) [26 (link)] and then used for ectopic expression of p53.

Ristic B., Sivaprakasam S., Narayanan M, & Ganapathy V. (2020). Hereditary hemochromatosis disrupts uric acid homeostasis and causes hyperuricemia via altered expression/activity of xanthine oxidase and ABCG2. Biochemical Journal, 477(8), 1499-1513.

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