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P1 nucleofection solution

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P1 Nucleofection solutions are a set of electroporation reagents designed for efficient delivery of nucleic acids into various cell types. The solutions enable transfection of DNA, RNA, or proteins into the cell nucleus, facilitating genetic manipulation and research applications.

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Lab products found in correlation

4d nucleofector x unit, by Lonza (6 mentions) Nrf2 cdna plasmids, by OriGene (2 mentions)

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Nucleofection (130 citations) Nucleofection solution (7 citations)

6 protocols using p1 nucleofection solution

1

Overexpression of CD38 and Nrf2 in CAMs

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Both CD38 cDNA (Catalog# MG204265) and Nrf2 cDNA plasmids (Catalog# MG226717) were purchased from OriGene Technologies. Transfection of plasmids was performed using a 4D Nucleofector X-Unit (Lonza, CA, USA) according to the manufacturer’s instructions, which was described in our recent studies [2 (link)]. Briefly, CAMs were trypsinized and centrifuged at 90×g for 10 minutes. The cell pellet was resuspended in 100 μL P1 Nucleofection solutions (Lonza) for Nucleofection (with the program code CM137). The program was chosen based on the fact that Nucleofection efficiency was over 80% as analyzed by flow cytometry using control GFP plasmids. For each Nucleofection sample, 2 μg plasmid DNA was added in 100 μL P1 Nucleofection solution. After Nucleofection, the cells were cultured in the DMEM medium for 24 hours and then were ready for treatment. The efficiency of CD38 cDNA and Nrf2 cDNA transfection was assessed by Western blot analyses [17 (link)].
Xu M., Li X.X., Wang L., Wang M., Zhang Y, & Li P.L. (2015). Contribution of Nrf2 to Atherogenic Phenotype Switching of Coronary Arterial Smooth Muscle Cells Lacking CD38 Gene. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 37(2), 432-444.
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2

Overexpression of DCTN2 and NOX1 in CAMs

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Both dynamitin (Dynactin 2: DCTN2) cDNA (Catalog# MC200162) and Nox1cDNA plasmids (Catalog# MG226022) were purchased from OriGene Technologies. DCTN2 cDNA plasmid contains a full-length DCTN2 gene (1653 bp) under a cytomegalovirus (CMV) promoter. Nox1cDNA plasmid contains a full-length Nox1 gene (1692 bp) under a CMV promoter. Transfection of plasmid was performed using a 4D Nucleofector X-Unit (Lonza, CA, USA) according to the manufacturer's instructions as we previously described [15 (link)]. Briefly, CAMs were tyrpsinized and centrifuged at 90×g for 10 minutes. The cell pellet was resuspended in 100 μL P1 Nucleofection solutions (Lonza) for Nucleofection (with the program code CM137). The program was chosen based on the fact that Nucleofection efficiency was over 80% as analyzed by flow cytometry using control GFP plasmids. For each Nucleofection sample, 2 μg plasmid DNA was added in 100 μL P1 Nucleofection solution. After Nucleofection, the cells were cultured in DMEM medium for 24 hours and then were ready for treatment. The efficiency of DCTN2cDNA and Nox1cDNA transfection was assessed by Western blot analyses.
Xu M., Li X.X., Chen Y., Pitzer A.L., Zhang Y, & Li P.L. (2014). Enhancement of dynein-mediated autophagosome trafficking and autophagy maturation by ROS in mouse coronary arterial myocytes. Journal of Cellular and Molecular Medicine, 18(11), 2165-2175.
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3

Overexpression of CD38 and Nrf2 in CAMs

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Both CD38 cDNA (Catalog# MG204265) and Nrf2 cDNA plasmids (Catalog# MG226717) were purchased from OriGene Technologies. Transfection of plasmids was performed using a 4D Nucleofector X-Unit (Lonza, CA, USA) according to the manufacturer’s instructions, which was described in our recent studies [2 (link)]. Briefly, CAMs were trypsinized and centrifuged at 90×g for 10 minutes. The cell pellet was resuspended in 100 μL P1 Nucleofection solutions (Lonza) for Nucleofection (with the program code CM137). The program was chosen based on the fact that Nucleofection efficiency was over 80% as analyzed by flow cytometry using control GFP plasmids. For each Nucleofection sample, 2 μg plasmid DNA was added in 100 μL P1 Nucleofection solution. After Nucleofection, the cells were cultured in the DMEM medium for 24 hours and then were ready for treatment. The efficiency of CD38 cDNA and Nrf2 cDNA transfection was assessed by Western blot analyses [17 (link)].
Xu M., Li X.X., Wang L., Wang M., Zhang Y, & Li P.L. (2015). Contribution of Nrf2 to Atherogenic Phenotype Switching of Coronary Arterial Smooth Muscle Cells Lacking CD38 Gene. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 37(2), 432-444.
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4

Overexpression of Dynactin2 and Nox1 in CAMs

Cited 1 time
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Both dynamitin (Dynactin 2: DCTN2) cDNA (Catalog no. MC200162) and Nox1cDNA plasmids (Catalog no. MG226022) were purchased from OriGene Technologies. DCTN2 cDNA plasmid contains a full-length DCTN2 gene (1653 bp) under a cytomegalovirus (CMV) promoter. Nox1cDNA plasmid contains a full-length Nox1 gene (1692 bp) under a CMV promoter. Transfection of plasmid was performed with a 4D Nucleofector X-Unit (Lonza, Anaheim, CA, USA) according to the manufacturer’s instructions as we previously described 15 (link). Briefly, CAMs were trypsinized and centrifuged at 90 × g for 10 min. The cell pellet was resuspended in 100 μl P1 Nucleofection solutions (Lonza) for Nucleofection (with the program code CM137). The program was chosen based on the fact that Nucleofection efficiency was over 80% as analysed by flow cytometry by using control GFP plasmids. For each Nucleofection sample, 2 μg plasmid DNA was added in 100 μl P1 Nucleofection solution. After Nucleofection, the cells were cultured in DMEM medium for 24 hrs and then were ready for treatment. The efficiency of DCTN2cDNA and Nox1cDNA transfection was assessed by Western blot analyses.
Xu M., Li X.X., Chen Y., Pitzer A.L., Zhang Y, & Li P.L. (2014). Enhancement of dynein-mediated autophagosome trafficking and autophagy maturation by ROS in mouse coronary arterial myocytes. Journal of Cellular and Molecular Medicine, 18(11), 2165-2175.
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5

Nucleofection of Nlrp3 shRNA Plasmids

Cited 1 time
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Transfection of shRNA plasmids was performed using a 4D Nucleofector X-Unit (Lonza, CA, USA) according to the manufacturer’s instructions [26 (link)]. The Nlrp3 shRNA plasmid was obtained from Origene. Briefly, MVECs were tyrpsinized and centrifuged at 80×g for 10 min. The cell pellet was resus-pended in 100 μL P1 Nucleofection solution (Lonza) for Nucleofection (with the program code CM137). The program was chosen based on the fact that Nucleofection efficiency was over 80 % as analyzed by flow cytometry using control GFP plasmids. For each Nucleofection sample, 2 μg plasmid DNA was added in 100 μL P1 Nucleofection solution. After Nucleofection, cells were cultured in DMEM medium for 24 h.
Chen Y., Wang L., Pitzer A.L., Li X., Li P.L, & Zhang Y. (2016). Contribution of redox-dependent activation of endothelial Nlrp3 inflammasomes to hyperglycemia-induced endothelial dysfunction. Journal of molecular medicine (Berlin, Germany), 94(12), 1335-1347.
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6

Transfection of cDNA Plasmids in CASMCs

Cited 1 time
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Transfection of cDNA plasmids was performed using a 4D Nucleofector X-Unit (Lonza, CA, USA) according to the manufacturer's instructions as we previously described [13 (link)]. ASM cDNA plasmid (pcDNA3.1/V5-His mASM) encoding a full-length Smpd1 gene under a cytomegalovirus (CMV) immediate early promoter (a gift from Prof. Dr. Erich Gulbins, Essen, Germany). Briefly, CASMCs were tyrpsinized and centrifuged at 90×g for 10 minutes. The cell pelletet was resuspended in 100 L P1 Nucleofection solution (Lonza) for Nucleofection (with the program code CM137). The program was chosen based on the fact that Nucleofection efficiency was over 80% as analyzed by flow cytometry using control GFP plasmids. For each Nucleofection sample, 2 μg plasmid DNA was added in 100 L P1 Nucleofection solution. After Nucleofection, cells were cultured in DMEM medium for 24 hours and then cells were ready for the treatments.
Li X., Xu M., Pitzer A.L., Xia M., Boini K.M., Li P.L, & Zhang Y. (2014). Control of Autophagy Maturation by Acid Sphingomyelinase in Mouse Coronary Arterial Smooth Muscle Cells: Protective Role in Atherosclerosis. Journal of molecular medicine (Berlin, Germany), 92(5), 473-485.
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