Hek293t

Manufactured by Biowest

Sourced in France

HEK293T is a cell line derived from human embryonic kidney cells. It is a commonly used host cell line for the expression and production of recombinant proteins, viral vectors, and other biological products.

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Lab products found in correlation

Hek293t, by American Type Culture Collection (3 mentions) Dulbecco modified eagle medium (dmem), by Biowest (3 mentions) Dmem high glucose, by Biowest (2 mentions) Mem non essential amino acid, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Roswell park memorial institute (rpmi), by Biowest (2 mentions) Fetal bovine serum (fbs), by Biowest (2 mentions) Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle medium, by Biowest (1 mentions) Elisa, by BioLegend (1 mentions) Hek293t, by Leibniz Institute DSMZ (1 mentions) Zeocin, by InvivoGen (1 mentions) Fugene hd transfection reagent, by Promega (1 mentions) Docetaxel, by Merck Group (1 mentions) Lncap, by Biowest (1 mentions) Rabbit anti parp1 h 250, by Santa Cruz Biotechnology (1 mentions) Goat anti actin 1 19, by Santa Cruz Biotechnology (1 mentions) F 12k nutrient mixture medium, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Biowest (1 mentions) Mouse anti ha 11 16b12, by Fortrea (1 mentions)

6 protocols using hek293t

Generating Engineered Cell Lines for Preclinical Studies

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Green fluorescent protein and firefly luciferase-positive AsPC-1 cells (AsPC-1 WT) were generated in-house as described previously by Schaefer et al.^{28 (link)} Luciferase expressing Raji cells were generated as described previously by Seitz et al.^{27 (link)} AsPC-1 WT secreting latent TGF-β (AsPC-1-TGF) were generated via transduction of AsPC-1 WT with lentiviral vectors (LVV) encoding human TGF-β (UniProtKB-A0A024R0P8-1). The plasmid encoding human TGF-β was kindly provided by Stefan Edelburg (Miltenyi Biotec, Bergisch-Gladbach, Germany). Secretion of TGF-β was confirmed by ELISA (BioLegend, catalog no.: 432907). Human embryonic kidney-293 T cells (HEK293T, DSMZ no: Acc635) were obtained from DSMZ.
AsPC-1 WT and AsPC-1-TGF were cultured in RPMI 1640 (Biowest catalog no.: L0501) supplemented with 2 mM glutamine (Lonza, catalog no.: BE17-605E) and 10% fetal bovine serum (EXIMUS Maximus FBS, Catus Biotech, catalog no.: BS-2020-500). HEK293T cells were cultured in Dulbecco’s modified Eagle medium (Biowest, catalog no.: L0104) supplemented with 10% FBS. Cell lines and primary human cells were cultured at 37°C and 5% CO₂.

Werchau N., Kotter B., Criado-Moronati E., Gosselink A., Cordes N., Lock D., Lennartz S., Kolbe C., Winter N., Teppert K., Engert F., Webster B., Mittelstaet J., Schaefer D., Mallmann P., Mallmann M.R., Ratiu D., Assenmacher M., Schaser T., von Bergwelt-Baildon M., Abramowski P, & Kaiser A.D. (2022). Combined targeting of soluble latent TGF-ß and a solid tumor-associated antigen with adapter CAR T cells. Oncoimmunology, 11(1), 2140534.

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Stable Overexpression of B4GalT4 Variants

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A375 and HEK293T cells obtained from the American Type Culture Collection were cultured in Dulbecco’s Minimum Eagle Medium (DMEM High Glucose, Biowest) under 5% CO₂ and 37 °C. The medium was supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, 100 μg/ml streptomycin and 4 mM L-glutamine. Stable transfections with pSelect plasmids for protein expression were performed using the FuGENE HD transfection reagent (Promega) according to the manufacturer’s protocol. Selection of stable transfectants overexpressing different B4GalT4 variants was done in the complete media supplemented by 400 μg/ml of zeocin (InvivoGen). Analysis of stable transfectants was done by Western blotting and indirect fluorescent staining. After the selection the cell culture was maintained with a two-fold reduced zeocin concentration.

Shauchuk A., Szulc B., Maszczak-Seneczko D., Wiertelak W., Skurska E, & Olczak M. (2020). N-glycosylation of the human β1,4-galactosyltransferase 4 is crucial for its activity and Golgi localization. Glycoconjugate Journal, 37(5), 577-588.

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Prostate Cancer Cell Lines Characterization

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HEK293T, human prostate adenocarcinoma LNCaP, Du145 and PC3 cells lines were purchased from the American Type Culture Collection, Manassas, VA, USA. The Docetaxel resistant cell lines Du145 and PC3 were developed as previously described [31 (link)]. HEK293T were cultured in DMEM (BioWest, Nuaillé, France), Docetaxel sentitive and resistant Du145 cells and LNCaP cells were cultured in RPMI-1640 (BioWest) and PC3 DS and PC3 DR in F12K nutrient mixture medium (Thermo Fisher Scientific, Waltham, MA, USA). Culture media were supplemented with 10% fetal bovine serum (FBS), 2 mM l-glutamine, 100 U of penicillin/ml, 100 μg of streptomycin/ml, and 0.1 mM non-essential amino acids (all from BioWest). Docetaxel resistant cell lines were maintained with 2.5 nM of Docetaxel (Sigma-Aldrich, St. Louis, MO). Antibody to PTOV1 was produced and purified as previously described [8 (link), 9 (link)]. Additional antibodies were obtained from: goat anti-actin (I-19) (Santa Cruz Biotechnology, Santa Cruz, CA); mouse anti-HA.11 (16B12) (Covance, MA); mouse anti-Cyclin B1(V152) (Abcam, Cambridge, MA); rabbit anti-PARP1 (H-250) (Santa Cruz Biotechnology). Lentiviral vectors carrying short-hairpin RNA (shRNA, TRCN0000143905, TRCN0000140104 and TRCN0000139737) to PTOV1 were obtained from Sigma.

Cánovas V., Puñal Y., Maggio V., Redondo E., Marín M., Mellado B., Olivan M., Lleonart M., Planas J., Morote J, & Paciucci R. (2017). Prostate Tumor Overexpressed-1 (PTOV1) promotes docetaxel-resistance and survival of castration resistant prostate cancer cells. Oncotarget, 8(35), 59165-59180.

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Culturing K562 and HEK293T Cell Lines

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K562 and HEK293T cell lines were obtained from the American Type Culture Collection (ATCC) and were used in experiments at their early passages. K562 cells were cultured in RPMI (Biowest) supplemented with 10% fetal bovine serum (FBS) (Biowest). HEK293T cells were cultured in DMEM supplemented with 4500 mg/L glucose (Biowest), 2 mM L-glutamine (Thermo Scientific), 1X MEM non-essential amino acids (Thermo Scientific), 1 mM sodium pyruvate, and 10% fetal bovine serum (FBS) (Biowest). All cell lines were tested for mycoplasma every 6 months.

Teo W.W., Cao X., Wu C.S., Tan H.K., Zhou Q., Gao C., Vanuytsel K., Kumar S.S., Murphy G.J., Yang H., Chai L, & Tenen D.G. (2022). Non-coding RNA LEVER sequestration of PRC2 can mediate long range gene regulation. Communications Biology, 5, 343.

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Cell Line Characterization and Cultivation

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SNU-398 (CRL-2233, RRID: CVCL_0077, Homo sapiens, male, Asian, 42-years-old, anaplastic HCC), PLC/PRF/5 (CRL-8024, RRID: CVCL_0485, Homo sapiens, hepatoma) and HEK 293T (CRL-11268, RRID: CVCL_0063, Homo sapiens, fetus, kidney) cell lines were purchased from the ATCC. Huh7 (JCRB0403, RRID: CVCL_0336, Homo sapiens, male, Asian, 57-years-old, differentiated hepatoma) was obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB). Cells were tested for Mycoplasma and characterized using STR profiling by the ATCC and JCRB, respectively. Cells were used within six months of resuscitation. Unless otherwise stated, HEK 293T, Huh7, and PLC/PRF/5 were grown in DMEM High Glucose (biowest) supplemented with 10% FBS (biowest) and SNU398 cells were cultured in RPMI-1640 medium (biowest) also supplemented with 10% FBS. Transfections were carried out in Opti-MEM (Gibco, Thermo Fisher Scientific) in combination with lipofectamine 2000 transfection reagent (Thermo Fisher Scientific). All the cell lines were cultured in a humidified incubator at 37°C, with 5% CO₂.

Bellido Molias F., Sim A., Leong K.W., An O., Song Y., Ng V.H., Lim M.W., Ying C., Teo J.X., Göke J, & Chen L. (2021). Antisense RNAs Influence Promoter Usage of Their Counterpart Sense Genes in Cancer. Cancer Research, 81(23), 5849-5861.

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Cell Line Maintenance and Validation

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K562 and HEK293T cell lines were obtained from the American Type Culture Collection (ATCC) and were used in experiments at their early passages. K562 cells were cultured in RPMI (Biowest) supplemented with 10% fetal bovine serum (FBS) (Biowest). HEK293T cells were cultured in DMEM supplemented with 4500 mg/L glucose (Biowest), 2mM L-glutamine (Thermo Scientific), 1X MEM non-essential amino acids (Thermo Scientific), 1mM sodium pyruvate, and 10% fetal bovine serum (FBS) (Biowest). All cell lines were tested for mycoplasma every six months.

Teo W.W., Cao X., Wu C., Tan H.K., Zhou Q., Yang H., Chai L., & Tenen D.G. (2020). RNA LEVER Mediates Long-Range Regulation of ε-globin by Keeping PRC2 in Check.

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