Polyethylenimine pei transfection reagent

Manufactured by Polysciences

Sourced in United States

Polyethylenimine (PEI) is a cationic polymer commonly used as a transfection reagent. It forms complexes with nucleic acids, facilitating their delivery into cells. PEI's ability to condense and protect nucleic acids makes it useful for various in vitro and in vivo applications.

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Lab products found in correlation

Dmem high glucose, by Euroclone (2 mentions) Gentamycin, by Thermo Fisher Scientific (1 mentions) Triton x 45, by Merck Group (1 mentions) Amphotericin b, by Thermo Fisher Scientific (1 mentions) Cell counting kit 8 cck 8, by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) Ex cell 293 medium, by Merck Group (1 mentions) Freestyle 293 expression medium, by Thermo Fisher Scientific (1 mentions)

4 protocols using polyethylenimine pei transfection reagent

Transient transfection of HEK 293T cells to evaluate Tp9 expression

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To evaluate the expression and secretion of Tp9, HEK 293T cells were transiently transfected with pCMV-Tp9AU1, pCMV-ΔspTp9AU1, pCMV-tPAspTp9AU1, pEF1α-tPA-Tp9-iresGFP, pCMV-Tp9spGFP, pEF1α-GFP (pWPI, Addgene), and pCMV-GFP (pEGFP-C1, negative control, Clontech) using Polyethylenimine (PEI) transfection reagent (Polysciences, Inc.). Briefly, cells were seeded at 3 × 10⁵ cells/well in 6-well plates and incubated overnight at 37°C/5% CO₂ in a humidified incubator. Cells were then incubated for 6 h with a transfection mix containing 3 μg plasmid DNA and PEI (ratio 1:2.5 DNA-PEI) in Dulbecco's modified essential medium (DMEM) high glucose (Euroclone) without serum. After incubation, the transfection mix was replaced by fresh medium EMEM, with 10% FBS, 100 IU/ml of penicillin, 100 μg/ml of streptomycin and 0.25 μg/ml of amphotericin B, and incubated for 24 h at 37°C/5% CO₂ in a humidified incubator. To test supernatant protein expression, the transfection solution was replaced with fresh DMEM/F12 (ratio 1:1) medium without FBS and incubated for 48 h at 37°C/5% CO₂ in a humidified incubator. Cell supernatant was then collected and analyzed by immunoblot.

Bastos R.G., Franceschi V., Tebaldi G., Connelley T., Morrison W.I., Knowles D.P., Donofrio G, & Fry L.M. (2019). Molecular and Antigenic Properties of Mammalian Cell-Expressed Theileria parva Antigen Tp9. Frontiers in Immunology, 10, 897.

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Reagents for Cell Culture Experiments

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Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), gentamycin, and amphotericin B were obtained from GIBCO-Invitrogen (Carlsbad, CA, USA). Dulbecco’s phosphate-buffered saline (PBS) was obtained from Hyclone (Logan, UT, USA). Cell Counting Kit-8 (CCK-8) and Triton X-45 were purchased from Sigma–Aldrich. Polyethylenimine (PEI) transfection reagent was obtained from Polysciences (Warrington, PA, USA). Tri-n-butyl phosphate (TnBP) and Triton X-100 were from Merck KGaA (Darmstadt, Germany).

Belem W.F., Liu C.H., Hu Y.T., Burnouf T, & Lin L.T. (2022). Validation of Viral Inactivation Protocols for Therapeutic Blood Products against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). Viruses, 14(11), 2419.

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Expression and Purification of hSARM1 Protein

Cited 1 time

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hSARM1 and mutants (residues 28–724 in PSF-CMV-AMP,
N-terminal His₆-tag, TEV-protease cleavage site, AVI-tag) (Figley et al., 2021 (link)) were expressed in
HEK293S cells (ATCC) and purified to homogeneity using a combination of IMAC
and SEC. The HEK293S cell experiments were performed in accordance with
Griffith University Research Ethics Committee approval 2020/246. The cells
(were grown in 50 % Freestyle 293 Expression Medium (Gibco) and 50 % Ex-Cell
293 Medium (Sigma) supplemented with 3 % L-glutamine in vented flasks at 90
rpm in an 80 % humidified, 8 % carbon dioxide atmosphere at 37°C.
When cells reached a density of 2 × 10⁶ cells/mL,
they were centrifuged at 500 × g for 10 min and
resuspended in 100 % Freestyle 293 Expression Medium to a density of 2.5
×10⁶ cells/mL. After resuspension the cells were
transfected with 3 μg/mL of plasmid DNA using Polyethylenimine
(PEI) transfection reagent (Polysciences) and growth was continued
overnight. On the next day, transfected cells were diluted 1:1 with Ex-Cell
293 Medium and valproic acid was added to a final concentration of 2.2 mM.
Growth was continued for an additional three days. Cells were harvested by
centrifugation at 1,500 × g for 10 min at 4°C
and stored at −80°C until used for purification.

Shi Y., Kerry P.S., Nanson J.D., Bosanac T., Sasaki Y., Krauss R., Saikot F.K., Adams S.E., Mosaiab T., Masic V., Mao X., Rose F., Vasquez E., Furrer M., Cunnea K., Brearley A., Gu W., Luo Z., Brillault L., Landsberg M.J., DiAntonio A., Kobe B., Milbrandt J., Hughes R.O, & Ve T. (2022). Structural basis of SARM1 activation, substrate recognition and inhibition by small molecules. Molecular cell, 82(9), 1643-1659.e10.

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Transfection of HEK 293T cells

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HEK 293 T cells were seeded into six well plates (3 × 10⁵ cells/well) and incubated at 37°C with 5% CO₂. When cells were sub-confluent, the culture medium was removed and the cells were transfected with pIgK-PPRV-H-gD₁₀₆, pINT₂-PPRV-H-gD₁₀₆, and pEGFP-C1 using Polyethylenimine (PEI) transfection reagent (Polysciences, Inc.). Briefly, 3 µg of DNA were mixed with 7.5 µg PEI (1 mg/ml) (ratio 1:2.5 DNA:PEI) in 200 µl of Dulbecco’s modified essential medium (DMEM) high glucose (Euroclone) without serum. After 15 min at room temperature, 800 µl of medium without serum were added, and the transfection solution was transferred to the cells (monolayer) and left for 6 h at 37°C with 5% CO₂, in a humidified incubator. The transfection mixture was then replaced with fresh medium EMEM, with 10% FBS, 100 IU/ml of penicillin, 100 µg/ml of streptomycin, and 0.25 µg/ml of amphotericin B, and incubated for 24 h at 37°C with 5% CO₂.

Macchi F., Rojas J.M., Verna A.E., Sevilla N., Franceschi V., Tebaldi G., Cavirani S., Martín V, & Donofrio G. (2018). Bovine Herpesvirus-4-Based Vector Delivering Peste des Petits Ruminants Virus Hemagglutinin ORF Induces both Neutralizing Antibodies and Cytotoxic T Cell Responses. Frontiers in Immunology, 9, 421.

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