Wild-type (WT) HeLa cells were cultured in fetal bovine serum (FBS) containing MEM media (Gibco) to approximately 80% confluency in T-225 flasks. On the day of transfection, the cells were lifted from the flasks using TryplyE express (Gibco), counted, and resuspended to 1.1e7 vc/mL. Select FLAG-tagged HLA mRNA was serially diluted 2-fold across 15 points in SE transfection buffer (Lonza) in 96-well v-bottom plates. WT HeLa cells were added to each well containing the mRNA at a concentration of 1.33e7 vc/mL. The mRNA/cell mixture was transferred to a 16-well Lonza 4D cuvette and electroporated according to the manufacturer’s protocol established for HeLa cells. Post-transfection, the cells were immediately placed into serum containing MEM growth media and seeded into rows of 384-well culture plates at a density of 2,500–5,000 cells/well, depending on experiment. Remaining transfected HeLa cells were seeded into separate 96-well plates specifically for fluorescence-activated cell sorting (FACS) expression testing. Plates were cultured for 18 h at 37°C and 5% CO2.
4d cuvette
Manufactured by Lonza
The 4D cuvette is a laboratory equipment designed for spectrophotometric measurements. It features a four-sided cuvette design that allows for 360-degree optical access, enabling comprehensive sample analysis. The 4D cuvette is suitable for a variety of applications where precise optical measurements are required.
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Lab products found in correlation
2 protocols using 4d cuvette
1
HeLa Cell Electroporation of FLAG-HLA mRNA
2
Efficient mRNA Transfection of Mammalian Cells
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On the day of transfection, target cells (HeLa or K562, depending on experiment) were counted, washed with 1x PBS and resuspended to 1.1e7vc/mL in SE (for HeLa cells) or SF (for K562 cells) transfection buffer (Lonza). Target antigen mRNA was serially diluted 2-fold across 15 points in SE or SF transfection buffer in 96-well v-bottom plates. Target cells were added to each well containing the mRNA at a concentration of 1.33e7vc/mL. The mRNA/cell mixture was transferred to a 16-well Lonza 4D cuvette and electroporated according to the manufacturer’s protocol established for target cell line. Post-transfection, the cells were immediately placed into MEM growth media containing serum and seeded into rows of 384-well culture plates at a density of 5,000–10,000cells/well, depending on experiment. Remaining transfected cells were seeded into separate 96-well plates for expression testing by flow cytometry. Plates were cultured for >16 hours at 37°C and 5% CO2.
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