HUVECs were cultured and maintained in EBM medium supplemented with EGM-MV Single Quots (Lonza) at 37°C in 5% CO2. For live imaging, HUVECs were cultured at 50,000 cells/coverslip on 10 µg/ml fibronectin-coated glass coverslips (except for experiments using polyacrylamide substrates) and medium was supplemented with 25 µM Hepes, pH 7.2, and 30 U/ml Oxyrase. For Aurora A inhibition studies, cells were treated with 40 nM Aurora A Inhibitor I (Selleck Chemicals) for 60 min before imaging. Transfection of cDNAs and shRNA was performed using a Nucleofector Device with solution kit V (Lonza), setting A-034, and experiments were performed 6–10 h later to allow time for protein expression and/or shRNA knockdown.
Nucleofector device with solution kit 5
Manufactured by Lonza
The Nucleofector Device with Solution Kit V is a laboratory instrument designed for the efficient delivery of nucleic acids, such as DNA and RNA, into a variety of cell types. It utilizes a proprietary electroporation technology to facilitate the transfection of cells. The device and its accompanying solution kit provide a standardized and optimized system for cell transfection, enabling researchers to perform their experiments with consistency and efficiency.
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Lab products found in correlation
2 protocols using nucleofector device with solution kit 5
1
Live Imaging of HUVECs on Fibronectin
2
Endothelial Cell Culture and Transfection
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HUVECs (ECs) were cultured and maintained in EBM medium supplemented with EGM-MV Single Quots (Lonza, Walkersville, MD, USA) and penicillin/streptomycin (RPI) at 37°C with 5% CO2. Transfection of siRNA and/or cDNA was performed using a Nucleofector Device with solution kit V (Lonza), setting A-034. ECs were cultured at 50,000 cells/coverslip for live-cell imaging and branching morphogenesis, or 500,000 cells/coverslip for wound-edge migration on either uncoated glass, 2D collagen-I, or 2D fibronectin-coated glass coverslips (see 2D substrate below). Experiments were performed 8-12 h after transfection to allow time for protein expression and/or siRNA knockdown. Before live-cell imaging, the exposed cell surface was supplemented with 25 µm Hepes, pH 7.2, diluted with 1× PBS (Lonza), to maintain physiological pH.
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