Cell-light Tubulin-GFP (Thermo Fisher Scientific) was used to label tubulin with green fluorescent protein (GFP) in live infected cells. Cell-light Tubulin-GFP is a modified baculovirus that encodes the human tubulin gene fused to the GFP. BSR cells were co-infected with CVS-PmCherry and Cell-light Tubulin-GFP. Live-cell time-lapse experiments were recovered with a Zeiss AxioObserver epifluorescence microscope (63× oil-immersion objective). Images were deconvoluated using the Huygens Imaging software (Scientific Volume Imaging). A blind deconvolution algorithm has been used.
Celllight tubulin gfp
Manufactured by Thermo Fisher Scientific
Sourced in United States
The CellLight Tubulin-GFP is a fluorescent protein-based labeling system that enables the visualization of the microtubule cytoskeleton in live cells. It functions by expressing a fusion protein consisting of the green fluorescent protein (GFP) and the tubulin protein, which then incorporates into the microtubules within the cell.
Automatically generated - may contain errors
Lab products found in correlation
Bacmam 2, by Thermo Fisher Scientific (2 mentions)
Sir dna reagent, by Cytoskeleton (2 mentions)
Axio observer, by Zeiss (1 mentions)
Nucblue, by Thermo Fisher Scientific (1 mentions)
Lsm 980, by Zeiss (1 mentions)
35 mm coverslip bottom dishes, by MatTek (1 mentions)
Las x software, by Leica (1 mentions)
Tcs sp5 dmi6000 cs confocal microscope, by Leica (1 mentions)
4 protocols using celllight tubulin gfp
1
Live Imaging of Tubulin Dynamics in Infected Cells
2
Visualizing Microtubule Dynamics in HeLa Cells
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HeLa cells were treated with CellLight Tubulin-GFP and BacMam 2.0 (Thermo Fisher Scientific) for 12 h before siRNA transfection. Transfection of siRNAs into HeLa cells at 1 nM concentration was carried out as described above. Nuclei were visualized by staining of DNA with SiR-DNA reagent (Cytoskeleton) (0.25 μM) for 6 h. Cells were cultured in CellView 3.5 cm glass-bottomed dishes (Greiner). Time-lapse images were obtained using a Leica TCS SP5 DMI6000 CS Confocal Microscope between 48 and 72 h post transfection.
3
Intracellular Trafficking of Nanomedicines
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Approximately 5 × 104 cells were seeded in 35 mm coverslip-bottom dishes (MatTek, Ashland, MA, USA) with 2 mL of medium and were incubated for 24 h. One day after seeding, the cells were concurrently dosed with GNPPEG-CY5-RGD at 1 nM and the IC-50 dose of either DTX, LNPDTX-1, or LNPDTX-2. Then, 16 h prior to imaging, the tubulin stain CellLight™ Tubulin-GFP (C10613; BacMam 2.0, ThermoFisher Scientific, Waltham, MA, USA) was added. Next, 24 h post-treatment, 4 drops of the live reagent (DAPI) NucBlue® (R37605; ThermoFisher Scientific, Waltham, MA, USA) was added per dish, followed by incubation for 20 min. Live cell imaging was performed using a 60X oil-immersion lens for confocal microscopy (Zeiss LSM 980, Carl Zeiss Microscopy GmbH, Jena, Germany).
4
Imaging Tubulin and DNA in siADAR1-transfected HeLa Cells
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Transfection of siRNAs (siADAR1-1) into HeLa cells at 1 nM concentration was carried out as described above. After incubation for 24 h, the culture medium was replaced with a fresh medium containing CellLight Tubulin-GFP and BacMam 2.0 (Thermo Fisher Scientific). Nuclei were stained with SiR-DNA reagent (Cytoskeleton) at 0.25 μM for 6 h. Cells were cultured on Ibidi μDish 3.5 cm. After 72 h, cells were fixed with 4% paraformaldehyde and soaked in Dulbecco’s PBS. Microscopic images were obtained by using a Leica TCS SP5 DMI6000 CS Confocal Microscope and LAS X software (Leica), equipped with ultraviolet 405 diode, Argon, DPS3561, and HeNe594 lasers. Fluorescent images were captured with a 40× lens with a 512 × 512 frame. For multicolor experiments, the following wavelength settings were used: Tubulin-GFP (Ex 488 nm/Em 498–630 nm) and SiR-DNA reagent (Ex 647 nm/Em 657–800 nm). Nuclear morphological analysis was performed using 4′,6-diamidino-2-phenylindole (DAPI)-stained HeLa cells.
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