Hek293 cell

Manufactured by Thermo Fisher Scientific

Sourced in United States, China, Switzerland, United Kingdom

HEK293 cells are a well-established immortalized cell line derived from human embryonic kidney cells. These cells are commonly used in various research applications, including gene expression studies, protein production, and virus propagation.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

HEK293 (267 citations) HEK293 Flp-In T-Rex cells (68 citations) HEK293 Freestyle cells (31 citations) HEK293-FT packaging cells (12 citations) Hek293 EBNA cells (10 citations) HEK293-6E cells (5 citations) T-Rex HEK293 Tet-O cells (3 citations) Flp-In™-293 (HEK293) cells (2 citations) HEK293 (293-H) cells (2 citations) HEK293 Flip-In T-REx cells (2 citations)

217 protocols using hek293 cell

Epigenetic Regulation by EZH2 Inhibitors

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

Pfeiffer were obtained from ATCC. HEK293 cells were obtained from Thermo Scientific. Pfeiffer cells, were cultured in RPMI 1460 (Life Technologies, 22400105). HEK293 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies, 11965118). Culture media were supplemented with 10% of FBS (Life Technologies, 26140129).
Antibodies for immuno-blotting: EZH2 (CST, #5246), Histone H3 (CST, #4499), Histone H3 Lysine 27 Methyl 2(CST, #9728), Histone H3 Lysine 27 Methyl 3(CST, #9733). Antibody Histone H3 Lysine 27 Methyl 1 (EMD Millipore, #07–448). Anti-rabbit secondary antibody stored in 50% glycerol (Jackson Immunoresearch Laboratories, Inc., 205–035-108). GSK 126 and EPZ-6438 were purchased from AdooQ and Xcess Biosciences, respectively. Compounds were dissolved to 10 mM in dimethyl sulfoxide (DMSO).

Baker T., Nerle S., Pritchard J., Zhao B., Rivera V.M., Garner A, & Gonzalvez F. (2015). Acquisition of a single EZH2 D1 domain mutation confers acquired resistance to EZH2-targeted inhibitors. Oncotarget, 6(32), 32646-32655.

+ Open protocol

+ Expand

Transfection of Cell Lines with siRNAs and Plasmids

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

All cell lines, including HEK293 and KGN cells, were purchased from Shanghai Jihe Biotechnology Co., LTD (Shanghai, China). The HEK293 cells were cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA). The KGN cells were cultured in DMEM/F12 medium (Gibco). All the cell lines were supplemented with 10% fetal bovine serum (Gibco, USA) and 1% penicillin-streptomycin at 37 °C and with 5% CO2 in a humidified atmosphere. CircLRRC8A siRNA (si-circLRRC8A), EIF4A3-specific siRNA, miR-125a-3p mimics, anti-miR-125a-3p, si-NFE2L1 and their negative controls siRNA (si-NC) were purchased from Gene Pharma (Shanghai, China). CircLRRC8A-specific over-expression plasmid and negative control plasmid were purchased from GeneChem (Shanghai, China). HEK293 cells and KGN cells were seeded in a six-well plate with a density of 1 × 10⁶ cells, incubated overnight, were then transfected with the related reagents using Lipofectamine 2000 (Thermo Fisher Scientific, USA) as the manufacturer’s instruction.

Xing J., Zhang M., Zhao S., Lu M., Lin L., Chen L., Gao W., Li W., Shang J., Zhou J, & Zhu X. (2023). EIF4A3-Induced Exosomal circLRRC8A Alleviates Granulosa Cells Senescence Via the miR-125a-3p/NFE2L1 axis. Stem Cell Reviews and Reports, 19(6), 1994-2012.

+ Open protocol

+ Expand

MTT Assay for Epsilon Toxin Cytotoxicity

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

To assess the cytotoxic effects of recombinant epsilon toxin, MTT (3-(4, 5-dimethyl thiazol-2yl)-2, 5-diphenyl tetrazolium bromide) assay was carried out. HEK293 cells (ATCC # CRL-1573), the human embryonic kidney cells, were obtained from Pasteur institute cell bank (Tehran, Iran). HEK293 cells were cultured in RPMI1640 (Gibco, USA) containing 1% penicillin/ streptomycin (Gibco, USA) and 10% fetal bovine serum (FBS) (Gibco, USA) at 37°C and 5% CO
₂. After reaching 80% confluency, HEK293 cells were detached using Trypsin/EDTA (Gibco, USA) and 8000 cells were harvested in each well of 96-well plate containing RPMI1640 containing 1% penicillin/streptomycin and 10% FBS. After overnight incubation at 37°C and 5% CO
₂, the cells treated with 0.97, 1.95, 3.90, 7.80, 15.62, 31.25, 62.50, 125, 250, 325, 500, 1000, 2000, 2500, 3000 and 4000 μg/mL epsilon toxin for 24h at 37°C and 5% CO
₂. After that, the medium was exchanged with the 200 μL RPMI1640 containing 20 μL MTT solution (5 mg/mL) (Sigma-Aldrich, USA) and incubated for 4 h at 37°C and 5% CO
₂. Then, the medium was replaced with 100 μL dimethyl sulfoxide (Sigma-Aldrich, USA), and the optical density of each well was recorded at 570 nm using a microtiter plate reader (Biorad, USA). Phosphate buffer saline (PBS) was tested as the negative control. All tests were triplicated and analyzed three times.

Chehreara R., Karizi S.Z., Hosseini H.M., Mirhosseini S.A., Shafiei M., Amani J, & Kazemi R. (2021). Cell death analysis of recombinant mature epsilon toxin on the kidney cell line. Iranian Journal of Microbiology, 13(6), 832-838.

+ Open protocol

+ Expand

Engineered Cell Lines for TRP Channel Studies

Cited 1 time

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

HEK-293 cells (ATCC; Rockville, MD) were engineered to stably over-express human TRPV3 (in pcDNA3.1 D V5/His-TOPO) and other human TRP channels (TRPA1, TRPM8, TRPV1, TRPV2, and TRPV4), as previously described (Deering-Rice et al., 2011 (link)). HEK-293 cells were grown in DMEM:F12 (Invitrogen; Carlsbad, CA) containing 5% FBS and 1X penicillin/streptomycin (Invitrogen; Carlsbad, CA). HEK-293 cells stably over-expressing the various TRP channels were maintained in DMEM:F12 supplemented with 5% FBS, 1X penicillin/streptomycin, and 350 μg/ml Geneticin (Invitrogen). Cells were sub-cultured using trypsin and plated into 1% gelatin-coated 96- or 384-well plates for calcium imaging experiments. Human immortalized keratinocytes, HaCaT cells, were provided by Dr. Douglas Grossman M.D., Ph.D. of the Department of Dermatology, University of Utah. HaCaT cells were cultured in DMEM containing 5% FBS and 1X penicillin/streptomycin/glutamine. Cells were sub-cultured using trypsin and plated into 96-well plates for experiments.

Deering-Rice C.E., Mitchell V.K., Romero E.G., Abdel Aziz M.H., Ryskamp D.A., Križaj D., Venkat R.G, & Reilly C.A. (2014). Drofenine: a 2-APB analog with improved selectivity for human TRPV3. Pharmacology Research & Perspectives, 2(5), e00062.

+ Open protocol

+ Expand

Role of ECL2 in GPR21 Activation

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

To investigate the role of ECL2 in the activation of GPR21, three GPR21 mutants were generated. The residues 169–178 (immersed region of ECL2), residues 179–188 (cap region of ECL2), and residues 169–187 (the whole ECL2) were replaced by a flexible GS linker (GGSGGS), respectively. Wild‐type GPR21 and GPR21 mutants were cloned into pCDNA3.1 vector and expressed in HEK293 cells (Invitrogen). Plasmids were transfected in HEK293 cells using lipofectamine 3000 (Invitrogen). cAMP accumulation was performed using HTRF cAMP kit (Cisbio Bioassays) following the manufacturer's instructions. In brief, HEK293 cells were seeded into 384‐well plates. Then, 5 μl of cAMP‐d2 reagent and 5 μl of cAMP Eu‐cryptate antibody were added to each well. After incubation at room temperature for 1 h, fluorescence was measured using a microplate reader (Envision 2105, PerkinElmer) with excitation at 330 nm and emission at 620 and 665 nm. cAMP accumulation was calculated from a standard dose–response curve using GraphPad Prism 8.0 (GraphPad Software). The cAMP levels of GPR21 mutants were normalized to that of wild‐type GPR21 (100% level).

Wong T., Gao W., Chen G., Qiu C., He G., Ye F., Wu Z., Zeng Z, & Du Y. (2023). Cryo‐EM structure of orphan G protein‐coupled receptor GPR21. MedComm, 4(1), e205.

+ Open protocol

+ Expand

Detailed Cell Culture Protocols for Diverse Tumor Models

Cited 1 time

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

B16F10, EMT6, and 4T1 were purchased from ATCC. 4T1-GFP-Luc (4T1-Luc) cells were generated by transfection of the 4T1 cell line with pGreenFire lentiviral vector (System Biosciences). Apigmented B16F10 cells were generated by deleting tyrosinase-related protein-2 (TRP2), henceforth B16F10-TRP2 knockout (KO) (18 (link)). MC38 cells were a gift from J. Schlom (National Cancer Institute). Tumor cells were cultured in Dulbecco’s modified Eagle’s medium (ATCC) supplemented with 10% FBS (Thermo Fisher Scientific). HEK293 cells purchased from Life Technologies were cultured in Freestyle medium (Life Technologies). Cells isolated from the spleen or blood of B6 mice were cultured in RPMI-1640 (ATCC) with 10% FBS, 20 mM HEPES, 1 mM sodium pyruvate, 0.05 mM β-mercaptoethanol (Life Technologies), 100 units/ml penicillin (Life Technologies), 100 μg/ml streptomycin (Life Technologies), 2 mM L-alanyl,-L-glutamine (Life Technologies), and 1x minimal essential medium (Corning) non-essential amino acids. CTLL-2 cells purchased from ATCC were cultured in the same medium with 10% T cell culture supplement with concanavalin A (T-STIM with ConA, Corning). All cells and cell assays were maintained at 37°C and 5% CO₂. All cell lines were tested for mycoplasma contamination, and none tested positive.

Momin N., Mehta N.K., Bennett N.R., Ma L., Palmeri J.R., Chinn M.M., Lutz E.A., Kang B., Irvine D.J., Spranger S, & Wittrup K.D. (2019). Anchoring of intratumorally administered cytokines to collagen safely potentiates systemic cancer immunotherapy. Science translational medicine, 11(498), eaaw2614.

+ Open protocol

+ Expand

Cell Culture Maintenance Protocol

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

Vero cells (ATCC number CCL-81) were maintained at 37°C/5 % CO₂ in minimal essential medium (Life Technologies) with 7 % heat-inactivated foetal bovine serum (FBS; Life Technologies), 1 % penicillin/streptomycin (PS; 5000 U.ml⁻¹ and 5000 μg.ml⁻¹; Life Technologies) and 1% Glutamine (Gln; 200 mmol.l⁻¹; Life Technologies). HEK-293 cells (ATCC number CCL-1573) were maintained at 37°C/ 5 % CO₂ in the same medium supplemented with 1 % non-essential amino acids (Life Technologies).

Mohamed Ali S., Vega-Rúa A., Driouich J.S., de Lamballerie X., Failloux A.B, & Nougairède A. (2018). Comparison of chikungunya viruses generated using infectious clone or the Infectious Subgenomic Amplicons (ISA) method in Aedes mosquitoes. PLoS ONE, 13(6), e0199494.

+ Open protocol

+ Expand

Allelic Regulation of Genetic Variants

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

DNA sequences (0.3kb) surrounding rs2275675 or rs10911339 were PCR amplified using genomic DNA from homozygous subjects for the C-allele (rs2275675) and the T-allele (rs10911339) using following primers:rs2275675, 5′-GGGGTACCGTAGAAAGAAAAGCAGAAC-3′ (forward) and 5′-GAAGATCTGAGACCTGCACCAATAAG-3′ (reverse); rs10911339, 5′-GGGGTACCGGTATGGGTGCCTAGC-3′ (forward) and 5′-GAAGATCTCCAGGTGTGCAGACTTC-3′ (reverse). The PCR products were digested using restriction enzymes, inserted into pGL3-promoter luciferase vector (Promega), and mutated to generate the respective other allele using Site-Directed Mutagenesis Kit (Stratagene). All construct inserts were sequenced for orientation and authenticity.
HEK-293 (human embryonic kidney cell line) and Raji B cells were purchased from ATCC. HEK-293 cells were seeded on 24-well plates at 2×10⁵ cells/well and transiently transfected using Lipofectamine 2000 (Life Technologies) with 1 μg rs2275675 vector (C or T), rs10911339 vector (C or T) or empty vector (pGL3-promoter) and 100 ng pRL-SV40 vector (Renilla luciferase) as an internal control. Raji cells were seeded on 24-well plates at 2×10⁶ cells/well and electroporated with 5μg luciferase constructs and 100 ng Renilla control vector using nucleofector (Amaxa). Luciferase activity in cell lysates was measured after 24 hours using a dual luciferase reporter assay (Promega).

Deng Y., Zhao J., Sakurai D., Sestak A.L., Osadchiy V., Langefeld C.D., Kaufman K.M., Kelly J.A., James J.A., Petri M.A., Bae S.C., Alarcón-Riquelme M.E., Alarcón G.S., Anaya J.M., Criswell L.A., Freedman B.I., Kamen D.L., Gilkeson G.S., Jacob C.O., Merrill J.T., Gaffney P.M., Sivils K.M., Niewold T.B., Ramsey-Goldman R., Reveille J.D., Scofield R.H., Stevens A.M., Boackle S.A., Vilá L.M., Sohn W I.I., Lee S., Chang D.M., Song Y.W., Vyse T.J., Harley J.B., Brown E.E., Edberg J.C., Kimberly R.P., Cantor R.M., Hahn B.H., Grossman J.M, & Tsao B.P. (2016). Decreased SMG7 expression associates with lupus-risk variants and elevated antinuclear antibody production. Annals of the rheumatic diseases, 75(11), 2007-2013.

+ Open protocol

+ Expand

Transient Transfection of Cell Lines

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

C6, HeLa, and HEK293 cells (all purchased from DSMZ, Braunschweig, Germany), as well as HEK293 cells stably transfected with human PS1 or PS1D246A mutant constructs (kindly provided by Dr. Jochen Walter, University of Bonn), were maintained in DMEM supplemented with 10% fetal bovine serum, and 100 units ml^‐1 penicillin/streptomycin. Cells were tested for mycoplasma by indirect DNA stain using Hoechst 33342 (Sigma‐Aldrich). The identity of the cells was verified by the provider by multiplex PCR of minisatellite markers for HeLa and HEK293 cells. The rat origin of C6 cells was verified by PCR by the provider. Plasmids were transiently transfected using Lipofectamine LTX (Thermo Fisher) according to the manufacturer's recommendations. For siRNA knockdown of BACE1 (Life Technologies, siRNA ID# 105154) and ADAM17 (Life Technologies, siRNA ID# 112004), HEK293 cells were transfected first with siRNA duplexes on day 1, followed by transfection of the pcDNA3.1‐mPD‐L1‐myc plasmid on day 2 using Lipofectamine 3000 (Life Technologies) for both transfections.

Kummer M.P., Ising C., Kummer C., Sarlus H., Griep A., Vieira‐Saecker A., Schwartz S., Halle A., Brückner M., Händler K., Schultze J.L., Beyer M., Latz E, & Heneka M.T. (2021). Microglial PD‐1 stimulation by astrocytic PD‐L1 suppresses neuroinflammation and Alzheimer’s disease pathology. The EMBO Journal, 40(24), e108662.

+ Open protocol

+ Expand

Isolation and Culture of Rat SCG Neurons

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

Experiments were performed on cultured SCG neurons from 4-weeks old male Wistar rats. Animal procedures were approved by the Universidad de Colima ethics and biosecurity committee. Briefly, rats were decapitated under anesthesia and the ganglia were removed and placed in a Ca²⁺-free Hank´s solution (37°C) containing papain (20 U/ml). Thereafter, papain was replaced by a mixture of collagenase I (1.6 mg/ml) and dispase II (5 mg/ml). Mechanically dissociated cells were suspended twice in DMEM, centrifuged and plated onto poly-L-lysine-coated glass chips. Cells were incubated at 37°C (5% CO₂) with DMEM supplemented with 10% of heat-inactivated fetal bovine serum (FBS). For other experiments, HEK-293 cells (Life Technologies, Carlsbad, CA) were grown in DMEM supplemented with 5% FBS in the incubator and passaged every 3 days. Cells were transiently co-transfected with rat Kv2.1 and GFP cDNA, with lipofectin according to the manufacture’s recommendations. The next day, cells were plated onto the glass chips and GFP-positive cells were recorded 8–24 hrs later on.

Angel-Chavez L.I., Acosta-Gómez E.I., Morales-Avalos M., Castro E, & Cruzblanca H. (2015). Forskolin Suppresses Delayed-Rectifier K+ Currents and Enhances Spike Frequency-Dependent Adaptation of Sympathetic Neurons. PLoS ONE, 10(5), e0126365.

+ 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!