Cells were seeded into 96-well cell-culture plates (#TCP011024, JET BIOFIL) with 1 × 105 cells/0.1 ml/well, After 24 h culture, the supernatants were discarded and the cells were rinsed twice with 100 μl of sterile PBS, then 100 μl of 2 μg/ml anti-hGPC3 scFv-SpyTag diluted with PBS containing 2% FBS was added to each well and incubated at room temperature (RT) for 15 min with gentle shaking. After washing with PBS, 100 μl of 1.5 μg/ml fluorescein isothiocyanate (FITC)-labeled goat anti-mouse IgG F(ab’)2 fragment (#115-095-072, Jackson ImmunoResearch Laboratories) was added and incubated at RT for 15 min. After the final wash, 100 μl of sterile PBS was added to each well and the results were observed; images were captured with an Olympus inverted fluorescence microscope (Olympus, Tokyo, Japan).
Tcp011024
Manufactured by Jet Biofil
TCP011024 is a laboratory equipment designed for general research and analytical applications. It serves as a core component in various scientific workflows, enabling efficient sample handling and processing. The product specifications and capabilities are provided by the manufacturer, and no further interpretation or extrapolation is made regarding its intended use.
Automatically generated - may contain errors
2 protocols using tcp011024
1
GPC3 Expression Visualization in Cells
2
Coverslip Fixation of Trypanosoma brucei
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For fixation on coverslips, 2 × 106 cells per specimen were used, and for fixation in solution 1 × 106 cells per specimen were used. The cells were pelleted by centrifugation for 5 min at 800 g, followed by two washes with PBS, or with PBS supplemented with 3.3 mM glucose (Penta, G01102) in the case of bloodstream T. brucei cells.
For fixation on coverslips, the cells were re-suspended in 50 µl of PBS and transferred onto a coverslip (High Precision Ø 12 mm No. 1.5H, Marienfeld) placed in a 24-well plate (Jet Biofil, TCP011024), to facilitate subsequent steps. Following a 5 min incubation, the cells were washed with PBS and fixed with 500 µl of a freshly prepared fixation solution containing 4% formaldehyde (Sigma, F8775) and 4% acrylamide (Sigma, A8887) in PBS. For fixation in solution, the cells were directly re-suspended in the fixation solution and transferred to a poly-L-lysine (Sigma, P4707) coated coverslip in a 24-well plate. In both cases, the cells were fixed overnight at room temperature, followed by a PBS wash.
For fixation on coverslips, the cells were re-suspended in 50 µl of PBS and transferred onto a coverslip (High Precision Ø 12 mm No. 1.5H, Marienfeld) placed in a 24-well plate (Jet Biofil, TCP011024), to facilitate subsequent steps. Following a 5 min incubation, the cells were washed with PBS and fixed with 500 µl of a freshly prepared fixation solution containing 4% formaldehyde (Sigma, F8775) and 4% acrylamide (Sigma, A8887) in PBS. For fixation in solution, the cells were directly re-suspended in the fixation solution and transferred to a poly-L-lysine (Sigma, P4707) coated coverslip in a 24-well plate. In both cases, the cells were fixed overnight at room temperature, followed by a PBS wash.
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