The human cervical carcinoma HeLa cells were obtained from the First Hospital of Lanzhou University and cultured in Dulbecco's modi ed Eagle's medium (DMEM, Minghai Biochem, Lanzhou, China) supplemented with 10% fetal bovine serum (Minghai Biochem, Lanzhou, China) at a culture temperature of 37℃, 5% CO 2 in incubator (Thermo, USA). DNA transfection was carried out using Exfect2000 transfection reagent (Vazyme, Nanjing, China) as a mediator according to the manufacturer's instruction. The pEGFP-N1-PCBP1 and non-targeting negative control pEGFP-N1 was purchased from Invitrogen (Invitrogen Life Technologies, CA, USA). To transfect HeLa cells with plasmid vector, cells were plated into either 60 mm dish or a 100 mm dish and allowed to adhere for 24 h. Exfect2000 transfection reagent was utilized for the transfection. After pEGFP-N1 or pEGFP-N1-PCBP1 transfection, cells were cultured for 5 h and then the medium was replaced with fresh medium supplemented with 10% fetal bovine serum. Cells were harvested 24-48 h after transfection.
Pegfp n1 pcbp1
Manufactured by Thermo Fisher Scientific
Sourced in United States
PEGFP-N1-PCBP1 is a plasmid vector that expresses a fusion protein of the human gene PCBP1 and the green fluorescent protein (GFP) from the jellyfish Aequorea victoria. The vector allows for expression of the PCBP1-GFP fusion protein in mammalian cell lines.
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2 protocols using pegfp n1 pcbp1
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HeLa Cell Transfection and Expression
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HeLa Cell Transfection and Expression
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The human cervical carcinoma HeLa cells were obtained from the First Hospital of Lanzhou University and cultured in Dulbecco's modi ed Eagle's medium (DMEM, Minghai Biochem, Lanzhou, China) supplemented with 10% fetal bovine serum (Minghai Biochem, Lanzhou, China) at a culture temperature of 37℃, 5% CO 2 in incubator (Thermo, USA). DNA transfection was carried out using Exfect2000 transfection reagent (Vazyme, Nanjing, China) as a mediator according to the manufacturer's instruction. The pEGFP-N1-PCBP1 and non-targeting negative control pEGFP-N1 was purchased from Invitrogen (Invitrogen Life Technologies, CA, USA). To transfect HeLa cells with plasmid vector, cells were plated into either 60 mm dish or a 100 mm dish and allowed to adhere for 24 h. Exfect2000 transfection reagent was utilized for the transfection. After pEGFP-N1 or pEGFP-N1-PCBP1 transfection, cells were cultured for 5 h and then the medium was replaced with fresh medium supplemented with 10% fetal bovine serum. Cells were harvested 24-48 h after transfection.
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