Expi293 suspension cells

Manufactured by Thermo Fisher Scientific

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Expi293 suspension cells are a cell line derived from human embryonic kidney cells (HEK293) that can be grown in suspension culture. These cells are designed for the high-yield production of recombinant proteins and are commonly used in protein expression and biopharmaceutical research and development.

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Lab products found in correlation

Expi293 expression medium, by Thermo Fisher Scientific (4 mentions) Sf 900 3 sfm, by Thermo Fisher Scientific (2 mentions) Expifectamine, by Thermo Fisher Scientific (2 mentions) Ampicillin, by Merck Group (1 mentions) Luria broth, by Thermo Fisher Scientific (1 mentions) Kanamycin, by Merck Group (1 mentions) Streptactin xt 4flow column, by IBA Lifesciences (1 mentions) Fectopro transfection reagent, by Polyplus Transfection (1 mentions) Protino ni nta column, by Macherey-Nagel (1 mentions) Esf 921 insect cell culture medium, by Expression Systems (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Nanodrop 2000, by Thermo Fisher Scientific (1 mentions) Pcdna3.4 vector, by Thermo Fisher Scientific (1 mentions) Histrap hp column, by GE Healthcare (1 mentions) Hitrap, by Cytiva (1 mentions)

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Expi293 cells (248 citations) Expi293 (54 citations)

6 protocols using expi293 suspension cells

Bacterial and Insect Cell Culture Protocols

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Bacterial cultures for plasmid amplification E. coli DH5α and for protein expression BL21(DE3)pLysS were grown in Luria Broth (Thermo Fisher) media supplemented with appropriate selection antibiotic ampicillin or kanamycin (Sigma) and maintained at 37 °C at ambient CO₂ with shaking at 250 rpm.
Baculovirus was generated in Spodoptera frugiperda (Sf9) cells maintained in Sf-900 III SFM (Gibco) supplemented with 10% FBS Superior (MERCK, Biochrom). For insect protein expression, Trichoplusia ni (High Five) cells grown in Insect-XPRESS Protein-free Insect Cell Medium with L-Gln (Lonza) were infected with baculovirus. Insect cells were maintained at 27 °C at ambient CO₂ with shaking at 120 rpm.
Expi293 suspension cells (Thermo Fisher) were maintained in Expi293 Expression Medium (Thermo Fisher) on a 120 rpm shaking platform at 37°C in a 5% CO₂ humidified incubator. Protein was produced by transient transfection using ExpiFectamine (Thermo Fisher).

Borowska M.T., Boughter C.T., Bunker J.J., Guthmiller J.J., Wilson P.C., Roux B., Bendelac A, & Adams E.J. (2023). Biochemical and biophysical characterization of natural polyreactivity in antibodies. Cell reports, 42(10), 113190.

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Expi293 Cell Suspension Culture Protocol

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Expi293 suspension cells (ThermoFisher A14527) were grown at 37°C, 8% CO₂, and 125 rpm shaking in Expi293 Expression Medium (ThermoFisher A1435102). Cells at a density of around 2.0 × 10⁶ mL⁻¹ were transfected with plasmids at 1.1 mg/L culture, using 3 mg/L of polyethylenimine (PEI) ‘MAX’ (Polysciences 24765, 1 mg/mL in PBS). Cells were grown at 37°C, 8% CO₂, and 125 rpm shaking for 48 hr, then harvested at 3000 g for 20 min, flash frozen in liquid nitrogen, and stored at −80°C, until they were used.

Ohashi Y., Tremel S., Masson G.R., McGinney L., Boulanger J., Rostislavleva K., Johnson C.M., Niewczas I., Clark J, & Williams R.L. (2020). Membrane characteristics tune activities of endosomal and autophagic human VPS34 complexes. eLife, 9, e58281.

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Production and Purification of SARS-CoV-2 Proteins

Cited 2 times

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The pcDNA3.1 plasmids encoding the SARS-CoV-2 S ectodomain (amino acids 1 to 1208) followed by a fold-on trimerization motif and tags (8×HisTag, StrepTag, and AviTag) and the RBD (amino acids 331 to 519), were kindly provided by Prof. Felix Rey (Structural Virology Research Unit, Institut Pasteur Paris) [5 (link)]. The pETM11 expression vector encoding the His-tagged SARS-CoV-2 N protein was kindly provided by Dr. Nicolas Escriou (Laboratoire d’Innovation: Vaccins, Institut Pasteur Paris) [5 (link),18 (link),19 (link)].
The trimeric S glycoprotein and its RBD fragment were produced by transient expression of exponentially growing Expi293 suspension cells (Thermo Fisher Scientific, Waltham, MA) using FectoPRO® transfection reagent (Polyplus) following the manufacturer’s guidelines. Recombinant proteins were purified by affinity chromatography using Strep-Tactin®XT 4flow column (IBA lifesciences) according to the manufacturer’s instructions. The His-tagged SARS-CoV-2 N protein was bacterially expressed in E. coli BL21 (DE3) and purified as a soluble dimeric protein by affinity purification using a Ni-NTA Protino column (Macherey Nagel). The eluates were analysed in 4–20% gradient precast protein gels (Nippon Genetics).

Sarrigeorgiou I., Moschandreou D., Dimitriadis A., Tsinti G., Sotiropoulou E., Ntoukaki E., Eliadis P., Backovic M., Labropoulou S., Escriou N., Pouliakis A., Giannopoulou G., Gaitanarou E., Lazaridis K., Mentis A., Mamalaki A., Grouzi E, & Lymberi P. (2022). Combined monitoring of IgG and IgA anti-Spike and anti-Receptor binding domain long term responses following BNT162b2 mRNA vaccination in Greek healthcare workers. PLOS ONE, 17(11), e0277827.

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Baculovirus Protein Expression in Insect Cells

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Baculovirus was generated in Spodoptera frugiperda (Sf9) cells maintained in Sf-900 III SFM (Gibco) supplemented with 10% FBS (Sigma) and GlutaMAX (Gibco). For insect protein expression, Trichoplusia ni (High5, Expression Systems) cells grown in ESF 921 Insect Cell Culture Medium (Expression Systems) were infected with baculovirus. Insect cells were maintained at 27°C at ambient CO₂ with shaking at 120rpm.
Expi293 suspension cells (Thermo Fisher) were maintained in Expi293 Expression Medium on a 120rpm shaking platform at 37°C in a 5% CO₂ humidified incubator. Protein was produced by transient transfection using ExpiFectamine (Thermo Fisher).

Yen M., Ren J., Liu Q., Glassman C.R., Sheahan T.P., Picton L.K., Moreira F.R., Rustagi A., Jude K.M., Zhao X., Blish C.A., Baric R.S., Su L.L, & Christopher Garcia K. (2022). Facile Discovery of Surrogate Cytokine Agonists. Cell, 185(8), 1414-1430.e19.

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Expi293 Cell Culture Protocol

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Proteins were expressed in Expi293 suspension cells (Thermo Fisher). Cells were grown in Expi293 Expression Medium (Thermo Fisher) at 37 °C with 8% CO₂.

Blythe E.E., Gates S.N., Deshaies R.J, & Martin A. (2019). Multisystem proteinopathy (MSP) mutations in VCP/p97 increase NPLOC4·UFD1L binding and substrate processing. Structure (London, England : 1993), 27(12), 1820-1829.e4.

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Optimization and Characterization of PEDV S1S2J Protein

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The S1S2J gene fragment (630-800aa) of spike protein gene sequences from the PEDV AJ1102 strain (GenBank accession no. AFQ37598.1) was optimized, artificially synthesized, and cloned into the pcDNA3.4 vector (Invitrogen, USA) to generate the S1S2J protein with a C-terminus mouse IgG2a Fc tag and a N-terminus 10 × His tag. After transient expression in Expi293 suspension cells (Thermo Fisher Scientific, USA), the S1S2J fusion protein was enriched using a HisTrap HP column (GE Healthcare, USA) according to the manufacturer’s guidelines.
To determine the epitopes, the gene fragment of PEDV S1S2J was truncated into three segments with 45 bp overlap between neighboring parts and cloned into the pcDNA3.4 vector. The plasmids expressing truncated proteins fused to the Fc part of mouse IgG2a at the C-terminus were transiently expressed in Expi293 suspension cells and enriched using a HiTrap Protein A HP affinity purification column. Protein concentration was ascertained using the A280 absorption value of Nanodrop2000 (Thermo Fisher Scientific, USA).

Huang N., Lang Q., Li L., Ge L, & Yang X. (2023). Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein. AMB Express, 13, 74.

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