Hek293f

Manufactured by American Type Culture Collection

Sourced in United States, China

HEK293F is a human embryonic kidney cell line commonly used in cell culture and research applications. The HEK293F cell line is a suspension-adapted version of the original HEK293 adherent cell line, which was derived from human embryonic kidney cells transformed with sheared human adenovirus 5 DNA.

Automatically generated - may contain errors

Lab products found in correlation

10 protocols using hek293f

Culturing and Transfecting Mammalian Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human embryonic kidney cells (HEK293F, HEK293T) and monkey kidney cells (COS-1) were purchased from ATCC (Manassas, VA) and maintained in DMEM supplemented with 10% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin in a 5% CO₂ atmosphere in a humidified incubator at 37 °C. HEK293F cells were seeded in 12-well plates and transfected with 1–2 μg plasmid DNA using Lipofectamine 3000 according to the manufacturer’s protocol. Stable clones were selected with geneticin (G418) for pcDNA based plasmids or zeocin (Invitrogen, San Diego, CA) for pSecTag 2B based plasmids and were analyzed for protein expression and secretion by western blotting. For O-fucosylation analysis, HEK293T cells were cultured in DMEM supplemented with 10% bovine calf serum in 6 cm cell culture dishes. 0.5–1 μg plasmid DNA was transfected using polyethylenimine (PEI).

Hubmacher D., Schneider M., Berardinelli S.J., Takeuchi H., Willard B., Reinhardt D.P., Haltiwanger R.S, & Apte S.S. (2017). Unusual life cycle and impact on microfibril assembly of ADAMTS17, a secreted metalloprotease mutated in genetic eye disease. Scientific Reports, 7, 41871.

+ Open protocol

+ Expand

Molecular Cloning and Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293F (HEK, human embryonic kidney cells) cells were purchased from ATCC. HIV-1 YU2 gp120 and HIV-1 YU2 gp160 plasmids were gifts from Joseph Sodroski. Modified human osteosarcoma cells (HOS.T4.R5) were a gift from Nathaniel Landau. HEK293T cells were purchased from ATCC. The DNA purification was conducted using the Promega Miniprep Kit (Cat. No. A1222). BamH1 (Cat. No. R0136S) and Nde1 (R0111S) restriction enzymes and DNA ligase (Cat. No. M0202S) were purchased from New England Biolabs for plasmid digestion ligation. Mutagenesis was conducted using the Quik Change II XL Site-Directed Mutagenesis Kit (Cat. No. 200522), and reaction was conducted using the PFU Ultra Polymerase (Cat. No. 600380-51) provided by Agilent Technologies. All other reagents for protein purification were purchased from Sigma–Aldrich unless otherwise specified. Biotinylated-Trp3 peptide was purchased from Scilight-Peptide, Inc. The integrity of the purified peptide was confirmed by mass spectrometry, observed mass of 1561.11 Da versus expected mass of 1559.82 Da (Supplementary Figure S9).

Parajuli B., Acharya K., Bach H.C., Parajuli B., Zhang S., Smith AB I.I.I., Abrams C.F, & Chaiken I. (2018). Restricted HIV-1 Env glycan engagement by lectin-reengineered DAVEI protein chimera is sufficient for lytic inactivation of the virus. The Biochemical journal, 475(5), 931-957.

+ Open protocol

+ Expand

MERS-CoV RBD Expression in Mammalian and Insect Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human embryonic kidney 293F cells (HEK 293F) were obtained from ATCC (Manassas, VA, USA). The Spodoptera frugiperda. 21 cells (sf21) were obtained from Invitrogen. SP2/0 myeloma cells were kept by the Henan Provincial Key Laboratory of Animal Immunology (Zhengzhou, China). HEK 293F cells were cultured in SMM 293-TII medium (Sino biological, Beijing, China), and SP2/0 cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Solarbio, Beijing, China) supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, USA). The sf21 cells were cultured in Sf-900 II SFM (serum free Insect Cell Culture Medium, Invitrogen, USA) DH10Bac and JM109 were purchased from Weidi Biotechnology Co. Ltd. (Shanghai, China). The eukaryotic expression vectors pFastBac1 (Invirogen, USA) and pcDNA3.1 were used to express the MERS-CoV RBD and the RBD truncations, respectively.

Wang P., Ding P., Wei Q., Liu H., Liu Y., Li Q., Xing Y., Li G., Zhou E, & Zhang G. (2021). Precise location of two novel linear epitopes on the receptor-binding domain surface of MERS-CoV spike protein recognized by two different monoclonal antibodies. International Journal of Biological Macromolecules, 195, 609-619.

+ Open protocol

+ Expand

Authenticated Cell Line Culturing Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293 and HEK293F cells were purchased from ATCC (CRL-1573) and Thermo Fisher Scientific (R79007), respectively. As HEK293 is on the International Cell Line Authentication Committee’s list of commonly misidentified cell lines, the cells used in this study were authenticated by short tandem repeat (STR) DNA profiling and tested negative for mycoplasma contamination. The culture medium was DMEM (4.5 g/L glucose, Corning 10013CV) supplemented with 100 μg/mL penicillin-streptomycin and 10% fetal bovine serum. Human NPE cells were purchased from ScienCell Research Laboratories (Cat. #6580), cultured in Epithelial Cell Medium (EpiCM, Cat. #4101) and authenticated by morphology. No mycoplasma contamination was found by DAPI staining.

Owji A.P., Yu K., Kittredge A., Wang J., Zhang Y, & Yang T. (2022). Bestrophin-2 and glutamine synthetase form a complex for glutamate release. Nature, 611(7934), 180-187.

+ Open protocol

+ Expand

Authenticated Cell Lines and Knockout Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Authenticated HEK293F, HEK293T, and HeLa cells were from ATCC and used at low passage; Sf9 cells were purchased from Thermo Fisher Scientific (Waltham, MA); Mouse embryonic fibroblasts from LAMP1/LAMP2 double knockout mice (Bandyopadhyay et al., 2008 (link); Eskelinen et al., 2004 (link)) were the generous gift of Dr. Paul Saftig (Christian-Albrechts-Universität Kiel, Germany). Mycoplasma contamination was monitored by DAPI staining.

Li J, & Pfeffer S.R. (2016). Lysosomal membrane glycoproteins bind cholesterol and contribute to lysosomal cholesterol export. eLife, 5, e21635.

+ Open protocol

+ Expand

Cell Line Acquisition and Culture

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293F, HEK293T and T47D were obtained from ATCC. The murine breast cancer cell line E0771 was obtained from CH3 BioSystems. Engineered cell lines Ddr1-KO E0771 (E0771-KO), Ddr1-KO, empty vector-reconstituted E0771 (E0771-EV) and Ddr1-KO, hDDR1-reconstituted E0771 (E0771-hDDR1) are described in our previous study.17 (link) The DDR1 overexpressing HEK293 (HEK293-DDR1) cell line was constructed by transducing HEK293T cells with a lenti-vector containing a human DDR1 (full length) gene construct. T47D was cultured in Roswell Park Memorial Institute (RPMI) 1640 media (catalog no. 10-040-CV, Corning) with 10% fetal bovine serum (FBS, catalog no. F0900-050, GenDEPOT). HEK293T and E0771 and its derivatives were cultured in Dulbecco’s Modified Eagle Medium (DMEM, catalog no. 10-013-CV, Corning) with 10% FBS.

Liu J., Chiang H.C., Xiong W., Laurent V., Griffiths S.C., Dülfer J., Deng H., Sun X., Yin Y.W., Li W., Audoly L.P., An Z., Schürpf T., Li R, & Zhang N. (2023). A highly selective humanized DDR1 mAb reverses immune exclusion by disrupting collagen fiber alignment in breast cancer. Journal for Immunotherapy of Cancer, 11(6), e006720.

+ Open protocol

+ Expand

Cell Culture Protocols for HEK 293T, HEK 293F, and TF-1

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK 293T, HEK 293F and TF-1 cells were obtained from the American Type Culture Collection (ATCC). HEK 293T and HEK 293F cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Grand Island, NY, USA) and FreeStyle™ 293 medium (Invitrogen, Carlsbad, CA, USA) respectively, supplemented with 10% FBS (Gibco) and 1% penicillin–streptomycin (10,000 U/mL) (Gibco). TF-1 cells were cultured in RPMI 1640 containing 20% FBS and 50 U/mL GM-CSF (Thermo Fisher Scientific).

Ma L., Zhu M., Li G., Gai J., Li Y., Gu H., Qiao P., Li X., Ji W., Zhao R., Wu Y, & Wan Y. (2022). Preclinical development of a long-acting trivalent bispecific nanobody targeting IL-5 for the treatment of eosinophilic asthma. Respiratory Research, 23, 316.

+ Open protocol

+ Expand

Cell Culture Conditions for Multiple Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK-293F, CHO-S, Raji, SK-OV-3 and Jurkat E6.1 cells were obtained from the American Type Culture Collection (ATCC). HEK-293F cells and CHO-S cells were cultured in FreeStyle™ 293 or FreeStyle™ CHO expression medium (Invitrogen, Carlsbad, CA, USA) respectively, supplemented with 1% Penicillin–Streptomycin (10,000 U/mL) (Invitrogen). Raji cells and SK-OV-3 cells highly expressing CD47 were grown in RPMI1640 containing 10% FBS (Gibco, GrandIsland, NY, USA) and 1% Penicillin–Streptomycin. Jurkat E6.1 cells were cultured in RPMI1640 supplemented with 10% heat-inactivated fetal calf serum (FCS) (Biological Industries, Kibbutz Beit Haemek, Israel), 1% Penicillin–Streptomycin and 2 mM L-glutamine (Gibco).

Ma L., Zhu M., Gai J., Li G., Chang Q., Qiao P., Cao L., Chen W., Zhang S, & Wan Y. (2020). Preclinical development of a novel CD47 nanobody with less toxicity and enhanced anti-cancer therapeutic potential. Journal of Nanobiotechnology, 18, 12.

+ Open protocol

+ Expand

Esophageal and Kidney Cell Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols

Shantou human embryonic esophageal (SHEE) cell line were obtained from professor Enmin Li of Shantou University and were cultured in DMEM containing 10% FBS at 37 °C in an atmosphere of 5% CO₂. EC cell lines (KYSE30, KYSE70, KYSE140, KYSE150, KYSE410, KYSE450, KYSE510) were obtained from the Chinese Academy of Sciences, HEK293T and HEK293F cells were purchased from American Type Culture Collection (ATCC; Manassas, VA). All cells were cytogenetically tested and authenticated before freezing. EC cells were maintained in RPMI-1640 medium with 10% FBS. HEK293T cells were maintained in DMEM medium supplemented with 10% FBS.

Wu W., Xu J., Gao D., Xie Z., Chen W., Li W., Yuan Q., Duan L., Zhang Y., Yang X., Chen Y., Dong Z., Liu K, & Jiang Y. (2023). TOPK promotes the growth of esophageal cancer in vitro and in vivo by enhancing YB1/eEF1A1 signal pathway. Cell Death & Disease, 14(6), 364.

+ Open protocol

+ Expand

Cultivation of HCC Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

We obtained HCC cell lines (PLC/PRF/5, HepG2‐2.15, Hep3B, SK‐hep1, HLF, and Huh‐7) and HEK293F cells from the American Type Culture Collection or China Center for Type Culture Collection (CCTCC, Wuhan, China). The cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 0.1 mg/mL streptomycin in a 5% CO₂ incubator at 37°C. All cell culture reagents were purchased from Gibco (Invitrogen).

Song J., Zhang X., Ge Q., Yuan C., Chu L., Liang H., Liao Z., Liu Q., Zhang Z, & Zhang B. (2018). CRISPR/Cas9‐mediated knockout of HBsAg inhibits proliferation and tumorigenicity of HBV‐positive hepatocellular carcinoma cells. Journal of Cellular Biochemistry, 119(10), 8419-8431.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!