For the myogenin promoter reporter assay, C2C12 cells were transfected with myogenin promoter luciferase reporter plasmids (100 ng), miRNA mimics (100 µM), and in-house modified SV40 promoter-driven Renilla luciferase vector (pSV40-R.Luc; Promega, Madison, WI, USA) as an internal control. For the 3' UTR reporter assay, C2C12 cells were transfected with 3' UTR reporter plasmids (100 ng) and miRNA mimics (100 µM). The miRNA mimics (RiboBio, Guangzhou, Guangdong, China) were transfected into C2C12 cells using K2 transfection reagent (Biontex Laboratories GmbH, München, Germany) following the manufacturer's instructions. PEI transfection reagent (Polyplus, Illkirch, France) was used for the transfection of plasmids. The luciferase activity was measured using a Lumat3 LB9508 luminometer (Berthold Technologies, Bad Wildbad, Germany).
Pei transfection reagent
Manufactured by Polyplus Transfection
Sourced in France
PEI transfection reagent is a cationic polymer that facilitates the delivery of nucleic acids, such as DNA or RNA, into cells. It is commonly used for transient gene expression and gene silencing experiments in cell culture. The reagent forms complexes with the nucleic acids, which are then taken up by the cells, allowing for efficient transfection.
Automatically generated - may contain errors
Lab products found in correlation
Psv40 rluc, by Promega (1 mentions)
Lumat3 lb9508 luminometer, by Berthold Technologies (1 mentions)
Mirna mimics, by Promega (1 mentions)
K2 transfection reagent, by Biontex (1 mentions)
Gl 183, by Beckman Coulter (1 mentions)
Fes172, by Beckman Coulter (1 mentions)
Anti kir2ds4, by Beckman Coulter (1 mentions)
Anti igm pe, by Southern Biotech (1 mentions)
Anti igg pe, by Southern Biotech (1 mentions)
Facscalibur, by BD (1 mentions)
Cellquest, by BD (1 mentions)
Flp in tm system, by Thermo Fisher Scientific (1 mentions)
3 protocols using pei transfection reagent
1
Myogenin Promoter and 3' UTR Luciferase Assay
2
Transient Transfection of HEK-293T Cells with KIR Alleles
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HEK‐293T cells were transiently transfected using the linear polyethylenimine derivative jetPEI transfection reagent (Polyplus, New York, NY) following the manufacturer's instruction. The pcDNA 3.1 plasmids used in the study code for: 2DL1*002, *003, *004, *012; 2DL2*001; 2DL3*002, *005; 2DS1*001, *002; 2DS2*001, 2DS4*001, and 2DS5*002. Forty‐eight hours after the transfection, the cells were tested with a panel of anti‐KIR2D mAb followed by anti‐IgG‐PE or anti‐IgM‐PE (Southern Biotechnology) by immunofluorescence and flow cytometric analysis (FACSCalibur and Cell Quest software, BD Biosciences). The mAbs used in these experiments were: 143211 (R&D Systems, Minneapolis, MN), 11PB6 (anti‐KIR2DL1/S1 and anti‐KIR2DL3 allotypes carrying E35 and R50, Miltenyi Biotec, Bergisch Gladbach, Germany), HP‐MA4 (anti‐KIR2DL1/S1/S3/S5, Biolegend, San Diego, CA), GL‐183 (anti‐KIR2DL2/L3/S2, Beckman Coulter), and FES172 (anti‐KIR2DS4, Beckman Coulter).
3
HEK293 Cell Transfection Optimization
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HEK 293 cells stably expressing the GIPR (Flp-ln TM HEK-GIPR) and the CCK2R (Flp-ln TM HEK-CCK2R) were obtained using the Flp-In TM system (Invitrogen). Alternatively, HEK293T transiently expressing the GIPR were used. All HEK293 derived cell lines cells were maintained in Dulbecco's Modified Eagle's medium supplemented with 10% of fetal bovine serum (FBS), in a humidified atmosphere at 95% air and 5% CO2. MIN-6-B1clone (kindly given by Doctor Jun-Ichi Miyazaki) was maintained in culture in Dulbecco's Modified Eagle's medium 25mM glucose supplemented with 15% of FBS, 71 µM 2-mercaptoethenol. Transfections were performed using polyethylenimine (PEI) transfection reagent (1mg/mL, pH 7.4) (Polyplus). Plasmids were diluted in DMEM without FBS (ratio DNA (µg) / PEI (µL) 1:3). The mixture was mixed for 15 sec on a vortex, incubated for 15 min at room temperature and then deposited on the cells.
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