Hek293t

Manufactured by Thermo Fisher Scientific

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About the product

HEK293T is a widely used human embryonic kidney cell line. It is a derivative of the HEK293 cell line and is commonly used in research and development for the production and study of recombinant proteins, viral vectors, and other biological products.

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Thermo Fisher Scientific offers various specialized HEK293T cell lines, including:

- **GeneBLAzer™ H2-CRE-bla HEK293T Cells**: These cells contain the human Histamine Receptor 2 (H2) integrated into the CellSensor™ CRE-bla HEK293T cell line.
- **CellSensor™ SIE-bla HEK293T Cell Line**: This line features a beta-lactamase reporter gene under the control of the SIE response element.
- **CellSensor™ ISRE-bla HEK293T Cell Line**: Responsive to interferon alpha, this cell line can be used to probe the JAK/STAT signaling pathway.
- **CellSensor™ AP-1-bla HEK293T Cell Line**: Contains a beta-lactamase reporter gene under the control of the AP-1 response element.

These specialized HEK293T cell lines are commercially available from Thermo Fisher Scientific. Pricing for these products varies depending on the specific cell line and region. For the most accurate and up-to-date pricing information, please consult Thermo Fisher Scientific's official website or contact their authorized distributors.

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Spelling variants of this product

HEK293T-Rex (23 citations) HEK293T/17 cells (14 citations) HEK293T T-REx™ cells (4 citations) Flp-In™ T-REx™ 293 (HEK293T) cells (2 citations) HEK293T packing cells (1 citations) HEK293T and HEK293F cells (1 citations) HEK293T FT cells (1 citations) HEK293T Piezo1−/− cells (1 citations) HEK293T and HeLa cells (1 citations) Flip-In HEK293T cells (1 citations)

The spelling variants listed below correspond to different ways the product may be referred to in scientific literature.
These variants have been automatically detected by our extraction engine, which groups similar formulations based on semantic similarity.

1 470 protocols using hek293t

Culturing and Transfecting Prostate Cancer Cell Lines

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The PCa cell lines (PC-3, DU145, C4-2, LNCaP, 22RV1 and VCaP) were purchased from ATCC (Manassas, VA, USA). The HEK293T, DU145 and VCaP cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% of FBS (Thermo Fisher Scientific, Waltham, MA, USA). PC-3, C4-2, LNCaP, and 22RV1 cell lines were cultured in RPMI 1640 medium supplemented with 10% of FBS. The cells were maintained in a 37 °C humidified incubator supplied with 5% CO₂.
Lentiviral sgRNA constructs as well as packaging vectors were transfected into HEK293T cells with Lipofectamine 2000 (Thermo Fisher Scientific, Cat# 11668500). Control or gene-specific siRNAs were transfected using Lipofectamine^® RNAiMAX (Thermo Fisher Scientific, Cat# 13778) according to the manufacturer’s instruction. Approximately 75~90% transfection efficiencies were routinely achieved. The sequence information for sgRNA and siRNA, and packaging vectors used for lentivirus transfection, are listed in Supplementary Tables S1–S3.

Yan Y., Shi L., Ma T., Wang L, & Huang H. (2025). SNP rs9364554 Modulates Androgen Receptor Binding and Drug Response in Prostate Cancer. Biomolecules, 15(1), 64.

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Cell Culture Conditions for Liver and Kidney Cell Lines

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All cell lines, including HEK293-T, Huh7, HepG2, Hep3B, MHCC97-H cells, were purchased from Shanghai Zhongqiao xinzhou Biotechnology company. The HEK293-T, Huh7, MHCC97-H cells were routinely cultured in Dulbecco’s Modified Eagle Medium (D MEM, Gibco, USA), supplemented with 10% fetal bovine serum (FBS, Gibco, USA), and THLE-2, Hep3B, HepG2 were routinely cultured in Minimum Essential Media (MEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA) 1% Gluta MAX (Gibco, USA) and 1% sodium pyruvate (Gibco, USA). All cells were incubated at 37°C in a humidified incubator with 5% CO₂. All cell lines were confirmed by short tandem repeat (STR) analysis recently.

Zhang M., Bai J., Yuan H., Duan X., Yu L., Li Y., Li K., Rile S., Wang X., Wang H., Liu P., Yan J, & Wang C. (2025). BRD1 deficiency affects SREBF1-related lipid metabolism through regulating H3K9ac/H3K9me3 transition to inhibit HCC progression. Cell Death & Disease, 16(1), 104.

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Activation of Immune Cells via Transfection

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HEK293T, A549, and RAW264.7 cells were purchased from the American Type Culture Collection (ATCC). HEK293T and A549 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO) containing 10% heat-inactivated fetal bovine serum (FBS) (GIBCO) and 1% penicillin/streptomycin solution (P/S) (Invitrogen). RAW264.7 cells were maintained in RPMI 1640 medium (GIBCO) containing 10% FBS and 1% P/S solution. Bone marrow-derived macrophages (BMDMs) were isolated from femurs and tibias of mice, and differentiated in the RPMI1640 medium with 10% FBS, 1% P/S, and 1% M-CSF-conditioned medium for 7 days as we described previously [56 (link)]. For mouse primary peritoneal macrophage (PM) preparation, the mice (male, 6–8 weeks) were intraperitoneally (i.p.) injected with 3% fluid thioglycollate medium (BD). Three days later, peritoneal exudate cells were collected and cultured for 1 h, the medium was replaced, and the adherent monolayer cells were PMs. All cells were cultured at 37°C in a 10% CO₂ atmosphere. To activate the cells, poly(I:C) and poly(dA:dT) were transfected into BMDMs, PMs, or HEK293T cells by Lipofectamine 2000. The ratio of transfection reagent over ligands was 2 (μL/μg). Additionally, IFN-β was directly added into the macrophage culture medium to stimulate the cells.

Qiao Z., Li D., Zhang F., Zhu J., Liu S., Bai X., Yao H., Chen Z., Yan Y., Xu X, & Ma F. (2025). USP5 inhibits anti-RNA viral innate immunity by deconjugating K48-linked unanchored and K63-linked anchored ubiquitin on IRF3. PLOS Pathogens, 21(1), e1012843.

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Cell Culture Conditions Standardization

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HeLa, HEK293T, A549 and H1299 cells were purchased from ATCC (American Type Culture Collection). HeLa and HEK293T cells were cultured in DMEM (Dulbecco’s Modified Eagle’s medium, Gibco) media. A549 and H1299 cells were grown with RPMI-1640 (Roswell Park Memorial Institute − 1640, Gibco) media. Both DMEM and RPMI-1640 media were supplemented with 1% streptomycin/penicillin (Thermo Fisher Scientific) and 10% foetal bovine serum (FBS, Gibco). These cells were maintained at 37 °C in an incubator with humidified atmosphere and 5% CO₂.

Cheng S., Qiu Z., Zhang Z., Li Y., Zhu Y., Zhou Y., Yang Y., Zhang Y., Yang D., Zhang Y., Liu H., Dai Z., Sun S.L, & Liu S. (2025). USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression. Cell Communication and Signaling : CCS, 23, 56.

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Cell Line Characterization and Drug Treatments

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All cell lines used in this study, including HCC1806, HCC1937, and HEK293T cells, were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA) and validated by short tandem repeat analysis, and these cell lines tested negative for mycoplasma contamination. HCC1806 and HCC1937 cells were cultured in RPMI 1640 medium supplemented with 5% FBS. HEK293T cells were cultured in DMEM (Thermo Fisher, Grand Island, USA) with 5% FBS at 37°C with 5% CO₂. Primary human umbilical vein endothelial cells (HUVECs) were maintained in an EGM-2 Bullet Kit (CC-3162, Lonza, USA). AS1842856 (Cat#HY-100596) and apatinib (Cat#HY-13342S) were purchased from MCE (New Jersey, USA).

Fang H., Liang H., Yang C., Jiang D., Luo Q., Cao W.M., Zhang H, & Chen C. (2025). STAMBPL1 activates the GRHL3/HIF1A/VEGFA axis through interaction with FOXO1 to promote angiogenesis in triple-negative breast cancer. eLife, 13, RP102433.

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Culturing Colorectal Cancer Cell Lines

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The colorectal cancer cell lines LoVo, SW-620, HCT-116, and the human embryonic kidney cell line HEK293T were obtained from the American Type Culture Collection (ATCC). LoVo, SW-620, and HEK293T cells were cultured in DMEM (12800; Gibco). FBS (10%, v/v; Gibco) and 1% penicillin/streptomycin (15140148; Gibco) were added to the DMEM. All cells were cultured at 37 °C in a 5% (v/v) CO2 incubator.
For CRC sphere cells, 1 × 10⁵ tumor cells were cultured on 6-well ultra-low adhesion culture plates (3471; Corning) with 2 mL of stem cell medium (DMEM/F12) (31330095; Gibco) with 1:50 B27(17504044; Gibco), 20 ng/mL FGF (450-33; PERROTECH), 20 ng/mL EGF (100-47; PERROTECH), 4 μg/mL heparin (CC101; Macgene), 100 μg/mL apo-transferrin (CC109; Macgene), and 1% penicillin/streptomycin as previously described.

Deng J., Li Y., Yin L., Liu S., Li Y., Liao W., Mu L., Luo X, & Qin J. (2025). Histone lactylation enhances GCLC expression and thus promotes chemoresistance of colorectal cancer stem cells through inhibiting ferroptosis. Cell Death & Disease, 16(1), 193.

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Establishing Stable Cell Lines for Cancer Research

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The HEK 293T and the colorectal cancer cell lines(RKO, CT26)used in this study were obtained from the Cell Bank of the Chinese Academy of Sciences (CASCB, Shanghai, China). The RKO cell line was cultured in DMEM (GIBCO, NY, USA) supplemented with 10% fetal bovine serum (GIBCO) and 1% penicillin-streptomycin (GIBCO). The HEK 293T and CT26 cell lines were cultured in RPMI-1640 (GIBCO) medium supplemented with 10% FBS and 1% penicillin-streptomycin. All cell lines were maintained in a 5% CO2 humidified incubator (Thermo Fisher Scientific, MA, USA) at 37 °C.
To establish stable cell lines, the lentiviral vectors for Mfsd2a overexpression and S100A14 knockdown plasmids were designed by GenePharma (Shanghai, China). Target cells were transfected with these lentiviral vectors, and stable transfectants were selected using puromycin. Verification was performed using qRT-PCR and Western Blotting. Detailed sequences are listed in Table S3.

Sun L., Li X., Xiao Y., Yu W., Chen X., Wang Z., Xia N., Chen X., Chen M., Zhu H., Li J., Wei J., Han S, & Pu L. (2025). Mfsd2a suppresses colorectal cancer progression and liver metastasis via the S100A14/STAT3 axis. Journal of Translational Medicine, 23, 59.

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Isolation and Culture of Mouse Embryonic Palatal Mesenchymal Cells

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Mouse embryonic palatal mesenchymal cells (MEPM) were isolated from embryonic palatal tissue in embryos of C57BL/6J mice at gestational day E13.5^{69 (link)}. In brief, mice were euthanized with CO₂ at E13.5. Paired palatal shelves were isolated and rinsed in PBS and subsequently in 100 µL trypsin-EDTA (0.05%) (Thermo Fisher, 25300062) for 5 min at 37 °C with frequent agitation. Trypsinization was stopped by adding 1 mL of DMEM/F-12 medium (Gibco, C11330500BT) supplemented with 10% fetal bovine serum (FBS) (Vivacell, C04001-500) and 1% penicillin-streptomycin (PS) (Beyotime, C0222). MEPM were subsequently seeded on large dishes and cultured overnight at 37 °C. The next day, cells were washed with PBS, and fresh media was added. HEK293T (ATCC, CRL-11268), HEPM (ATCC, CRL-1486), and MC3T3-E1 (ATCC, CRL-2593) cell lines were obtained from the American Type Culture Collection. HEK293T cells were cultured in DMEM high glucose medium (Gibco, C11995500BT) and HEPM, MC3T3-E1 cells in α-MEM (Gibco, C12440500BT) supplemented with 10% FBS and 1% PS at 37 °C and 5% CO₂ in a humidified incubator. The culture medium was changed every three days. MS023 (MCE, HY-19615), or an equal volume of DMSO (Sigma, BCCD8942) vehicle, was added to cells for 24 h at a final concentration of 30 nM.

Meng L., Jiang Y., You J., Chen Y., Guo S., Chen L, & Ma J. (2025). PRMT1-methylated MSX1 phase separates to control palate development. Nature Communications, 16, 949.

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Lentiviral Transduction of BMSCs

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Human embryonic kidney 293T cells (HEK293T), serving as host cells for lentivirus packaging, were obtained from Genechem Co., Ltd. (Shanghai, China). Prior to the experiment, these cells were cultured in a 10 cm dish for two to three days until they reached 90–95% confluency. For lentivirus production, recombinant viral plasmids pLV-EGFP: T2A: Puro-U6 > Nol3(encoding full-length ARC protein, Genechem, Shanghai, China, vectorID:20221201), pLV-EGFP: F2A: Puro-U6 > Nol3_shRNA(OBiO Technology, Shanghai, China, vectorID: Y18978), control vector pLV-EGFP: F2A: Puro-U6 > NC2_shRNA(OBiO Technology, Shanghai, China, vectorID: GL427NC2), and packaging plasmids (including pLP1, pLP2, and pLP/VSVG) were transfected into HEK293T cells using Lipofectamine 2000(Thermo Fisher scientific, Massachusetts, USA, Cat#11668019).
At 48 h post-transfection, lentiviral particles were harvested from the supernatant of HEK293T cells and concentrated using a 0.45 μm filter. The concentrated virus solution was then added to BMSCs cultures for infection experiments at various multiplicities of infection (ranging from 0 to 100). Transfection efficiency was assessed 72 h post-infection using fluorescence microscopy and real-time quantitative PCR (RT-qPCR).

Shi J., Wen J, & Hu L. (2025). 17β-estradiol promotes osteogenic differentiation of BMSCs by regulating mitophagy through ARC. Journal of Orthopaedic Surgery and Research, 20, 35.

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Cell Line Culture Protocols

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All cell lines in this study were purchased from American Type Culture Collection (ATCC, USA). The human CRC cell lines HT-29, LS174T and Colo320 were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Bio Basic Cat# E600028), and HCT116, SW48, SW620 were cultured with Dulbecco’s Modified Eagle Medium (DMEM) (Gibco Cat# C11965500BT; the human embryonic kidney cell line HEK-293T, human breast cancer cell line MCF-7, gastric cancer cell line SNU-1 and pancreatic cancer cell line PANC-1 were cultured in DMEM. All the culture mediums were supplemented with 10% fetal bovine serum (FBS, Procell Cat# 164210), penicillin (100 U/mL) and streptomycin (100 µg/mL). All cell lines were cultured at 37 °C in a humidified atmosphere of 5% CO₂.

Zhou W., Sheng Y., Hu D., An Y., Yang M., Wang W., Basnet S., Yan J., Zhang S., Liu Q., Li Y., Tan Y., Gao J., Sun K, & Du C. (2025). Proteasome inhibition induces DNA methylation alteration by attenuating the synthesis of DNA methyltransferase 1 and 3B in colorectal cancer. Scientific Reports, 15, 8534.

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