Plasmids containing human WT p46Shc and deletion-mutant p46ShcδSH2 were constructed and confirmed.20 (link) The SH2 domain was deleted using forward primer 5’-GAGCTGCTCAGCCATGGACAC-3’ and reverse primer 5’-GAGCGGAAACTGTACCTCGAGT-3’, and EX-H9090-M62-p46Shcδ (329–425) SH2 was constructed. Primary hepatocytes were transfected with 2 μg DNA per 12 wells or 3 μg DNA per 6 wells for approximately 24 hours using the jetPEI-Hepatocyte DNA transfection reagent (89129-944; Polyplus) following the manufacturer’s instruction. BAX small interfering RNA (siRNA) (sc-29212) and control siRNA (sc-37007) were from Santa Cruz Biotechnology, Inc, and the transfection was performed using Lipofectamine 3000 Transfection Reagent (L3000001; Invitrogen) according to the manufacturer’s instructions. Briefly, VL-17A cells were seeded on a collagen-coated, 8-well, chambered glass slide and incubated with 10 pm BAX or control siRNA and 1 μL Lipofectamine 3000 reagent in serum-free medium for 24 hours.
Jetpei hepatocyte dna transfection reagent
Manufactured by Polyplus Transfection
Sourced in United States
JetPEI-Hepatocyte is a DNA transfection reagent specifically designed for the efficient delivery of genetic material into hepatocytes. It serves as a tool for researchers studying gene expression, protein production, and other applications in liver cell models.
Automatically generated - may contain errors
2 protocols using jetpei hepatocyte dna transfection reagent
1
Plasmid construction and transfection of primary hepatocytes
2
Transfection of Immortalized Rat Hepatocytes
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BRL cells, an immortalized normal rat hepatocyte line obtained from the cell bank of Academia Sinica, Shanghai, China, were seeded in 30-ml plastic flasks at 1x106 cells/well and cultivated in DMEM supplemented with 10% fetal bovine serum (FBS) in a humidified incubator containing 5% CO2 at 37°C. When cell density reached 60–70% confluence, transfection was performed according to the manufacturer’s instructions (Polyplus Transfection, New York, NY, USA). BRL cells were separately transfected with plasmid pcDNA3-rIL-10, pcDNA3 and normal saline for 24 h using JetPEI™-Hepatocyte DNA transfection reagent (Polyplus Transfection, New York, NY, USA). The transfection system contained 5 μg DNA and 16 μl JetPEI-Hepatocyte per flask.
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