A PRL-1 (human protein tyrosine phosphatase type 4A, member 1 [PTP4A1]) plasmid was purchased (Origene, Rockville, MD, USA). The CMV6-AC vector containing the PTP4A1 gene was digested with the restriction enzymes Sgf1 and Mlu1 and included a GFP reporter gene and neomycin. A lentiviral vector including the PRL-1 gene was acquired from SeouLin Bioscience (Seongnam, Republic of Korea). A pLenti-RSV-EF1a vector containing PRL-1 was constructed as a tagged protein with C-terminal GFP and puromycin. To generate a control group, we used the AMAXA pCMV-GFP vector (Lonza, Basel, Switzerland) containing kanamycin, and maxGFP was digested with the restriction enzymes Kpn1 and Bgl2.
Amaxa pcmv gfp vector
Manufactured by Lonza
Sourced in Switzerland
The AMAXA pCMV-GFP vector is a plasmid DNA construct that contains the green fluorescent protein (GFP) gene under the control of the cytomegalovirus (CMV) promoter. The GFP gene allows for the visualization of transfected cells.
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Lab products found in correlation
5 protocols using amaxa pcmv gfp vector
1
Plasmid Transfection for Protein Expression
2
PRL-1 Overexpression in Mesenchymal Stem Cells
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The PRL-1 plasmid (human protein tyrosine phosphatase type 4 A, member 1; PTP4A1) was purchased (Origene Inc., Rockville, MD, USA) and used to induce the overexpression of the PRL-1 gene. The CMV6-AC vector containing PRL-1, the GFP reporter gene, CMV promoters, and the antibiotic neomycin was digested with Sgf1 and Mlu1 restriction enzymes. The PRL-1 lentiviral plasmid was purchased from SeouLin Bioscience (Seongnam, Korea). The pLenti-RSV-EF1α vector including PRL-1 was constructed with a C-terminal GFP as well as the antibiotic puromycin. The AMAXA pCMV-GFP vector was obtained from Lonza (Basel, Switzerland). The resulting plasmid was confirmed by DNA sequencing. Naïve PD-MSCs (6 × 104 cells/cm2) were harvested and transfected by using an AMAXA system with a Human MSC Nucleofector Kit (Lonza) and lentiviral vector (SeouLin Bioscience). After transfections by each system, cells generated using AMAXA were selected by using 1.5 mg/ml neomycin. In addition, cells generated using lentiviral vector were selected by using 2 μg/ml puromycin for 7 days. We changed the medium every other day and observed changes in cell morphology. Cells were maintained at 37 °C in a humidified atmosphere containing 5% CO2.
3
Overexpression of PRL-1 in PD-MSCs
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PRL-1 plasmid (human protein tyrosine phosphatase type 4 A, member 1; PTP4A1) was purchased (Origene Inc., Rockville, MD, USA) and used to induce the overexpression of the PRL-1 gene. CMV6-AC vector containing PRL-1, the GFP reporter gene, CMV promoters and the antibiotic neomycin was digested with Sgf1 and Mlu1 restriction enzymes. PRL-1 lentiviral plasmid was purchased (SeouLin Bioscience, Seongnam, Korea). pLenti-RSV-EF1α vector including PRL-1 was constructed with a C-terminal GFP as well as the antibiotic puromycin. AMAXA pCMV-GFP vector was contained (Lonza, Basel, Switzerland). The resulting plasmid was confirmed by DNA sequencing. Naïve PD-MSCs (6 × 10 4 cells/cm 2 ) were harvested and transfected by using an AMAXA system with a Human MSC Nucleofector Kit (Lonza) and lentiviral vector (SeouLin Bioscience). After transfections by each system, cells generated using AMAXA were selected by 1.5 mg/ml neomycin. In addition, cells generated using lentiviral vector were selected by 2 µg/ml puromycin for 7 days. We changed the medium every other day and observed changes in cell morphology. Cells were maintained at 37 in a humidified atmosphere containing 5% CO 2 .
4
Overexpression of PRL-1 Gene in Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The PRL-1 plasmid (human protein tyrosine phosphatase type 4 A, member 1; PTP4A1) was purchased (Origene Inc., Rockville, MD, USA) and used to induce the overexpression of the PRL-1 gene. The CMV6-AC vector containing PRL-1, the GFP reporter gene, CMV promoters and the antibiotic neomycin was digested with Sgf1 and Mlu1 restriction enzymes. The PRL-1 lentiviral plasmid was purchased from SeouLin Bioscience (Seongnam, Korea). The pLenti-RSV-EF1α vector including PRL-1 was constructed with a Cterminal GFP as well as the antibiotic puromycin. The AMAXA pCMV-GFP vector was obtained from Lonza (Basel, Switzerland). The resulting plasmid was con rmed by DNA sequencing. Naïve PD-MSCs (6x10 4 cells/cm 2 ) were harvested and transfected by using an AMAXA system with a Human MSC Nucleofector Kit (Lonza) and lentiviral vector (SeouLin Bioscience). After transfections by each system, cells generated using AMAXA were selected by using 1.5 mg/ml neomycin. In addition, cells generated using lentiviral vector were selected by using 2 µg/ml puromycin for 7 days. We changed the medium every other day and observed changes in cell morphology. Cells were maintained at 37 ℃ in a humidi ed atmosphere containing 5% CO 2 .
5
Overexpression of PRL-1 Gene in Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The PRL-1 plasmid (human protein tyrosine phosphatase type 4 A, member 1; PTP4A1) was purchased (Origene Inc., Rockville, MD, USA) and used to induce the overexpression of the PRL-1 gene. The CMV6-AC vector containing PRL-1, the GFP reporter gene, CMV promoters and the antibiotic neomycin was digested with Sgf1 and Mlu1 restriction enzymes. The PRL-1 lentiviral plasmid was purchased from SeouLin Bioscience (Seongnam, Korea). The pLenti-RSV-EF1α vector including PRL-1 was constructed with a Cterminal GFP as well as the antibiotic puromycin. The AMAXA pCMV-GFP vector was obtained from Lonza (Basel, Switzerland). The resulting plasmid was con rmed by DNA sequencing. Naïve PD-MSCs (6x10 4 cells/cm 2 ) were harvested and transfected by using an AMAXA system with a Human MSC Nucleofector Kit (Lonza) and lentiviral vector (SeouLin Bioscience). After transfections by each system, cells generated using AMAXA were selected by using 1.5 mg/ml neomycin. In addition, cells generated using lentiviral vector were selected by using 2 µg/ml puromycin for 7 days. We changed the medium every other day and observed changes in cell morphology. Cells were maintained at 37 ℃ in a humidi ed atmosphere containing 5% CO 2 .
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