At 48 h after trans-fection, MPMCs were exposed to 10 mM EdU (Guangzhou RiboBio Co., Ltd., Guangzhou, China) for 24 h at 37°C. The cultured cells were then fixed with 4% paraformalde-hyde (PFA) for 30 min at room temperature and permeabilized with 0.5% Triton X-100. Next, the 1X Apollo reaction cocktail (Guangzhou RiboBio Co., Ltd.) was added to the cells and incubated for 30 min at room temperature. The cells were then stained using a Cell-Light™ EdU Apollo®488 in vitro imaging kit (Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. EdU-stained cells were observed using a fluorescence microscope (magnification, ×400; Nikon Corporation, Tokyo, Japan).
Cell light edu apollo 488 in vitro imaging kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Cell-Light™ EdU Apollo® 488 In Vitro Imaging kit is a reagent kit that enables the detection and visualization of proliferating cells in in vitro cell culture models. It utilizes the EdU (5-ethynyl-2'-deoxyuridine) labeling method to mark cells undergoing DNA synthesis, which is then visualized using a fluorescent azide dye.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microscope, by Nikon (3 mentions)
Apollo reaction cocktail, by RiboBio (1 mentions)
5 ethynyl 2 deoxyuridine (edu), by Nikon (1 mentions)
5 ethynyl 2 deoxyuridine (edu), by Thermo Fisher Scientific (1 mentions)
Goat serum, by Thermo Fisher Scientific (1 mentions)
Ix53 fluorescent microscope, by Olympus (1 mentions)
Pannoramic midi digital slide scanner, by 3DHISTECH (1 mentions)
6 protocols using cell light edu apollo 488 in vitro imaging kit
1
EdU Incorporation Assay for Proliferation
2
Cell Proliferation Assay using EdU
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Transfected cells (5×104 cells/well) were inoculated into 96-well plates and incubated with 20 µM EdU (Thermo Fisher Scientific, Inc.) at 37°C for 2 h. Subsequently, cells were fixed and permeated using 4% paraformaldehyde for 30 min and 0.5% Triton X-100 for 10 min at room temperature, respectively. The cells were stained with Cell-Light™ EdU Apollo® 488 In Vitro Imaging kit (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. Cells were observed using a fluorescence microscope (magnification, ×200; Nikon Corporation).
3
EdU Proliferation Assay Protocol
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The cells were seeded at ~1×104 cells per well. Following cultivation for 24 h, 20 µM EdU was added to the culture medium for 8 h at 37°C. The cultured cells were then fixed with 4% paraformaldehyde for 20 min. Triton X-100 (0.2%) was used to permeabilize the nuclear membrane, and PBS containing 10% goat serum (Gibco; Thermo Fisher Scientific, Inc.) was utilized for blocking for 1 h at room temperature. Finally, the cells were then stained using a Cell-Light™ EdU Apollo®488 In Vitro imaging kit (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol and images were obtained using a fluorescence microscope (Nikon Corporation, Tokyo Japan).
4
Evaluating miR-381 Effects on HUVEC Proliferation
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The EdU assay was performed to evaluate the effects of miR-381 on cell proliferation. HUVECs were seeded (1×104 cells/well) into 96-well plates 12 h prior to transfection. OX-LDL (80 µg/ml) was added to the cells 24 h following transfection and incubated for another 48 h at 37°C. EdU (100 µl) was added and cells were incubated at 37°C for another 8 h. The cells were fixed with 4% paraformaldehyde at room temperature for 20 min, permeabilized with Triton X-100, and blocked with PBS containing 10% goat serum for 1 h at 25°C. Cells were stained by Cell-Light™ EdU Apollo® 488 In Vitro Imaging kit (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. The images were captured under an IX53 fluorescent microscope (magnification, ×200; Olympus Corporation). EdU-positive cells were counted in three randomly selected fields.
6
EdU Assay for Cellular Proliferation
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For the EdU assay, si-KLF4/BMI1, control siRNA, LV-miR-135a or the corresponding NC were transfected, and PTC-209 or DMSO was added to Saos-2 cells. Then, the cells were cultured in the supplement of EdU for 8 h, and the cells were fixed with 4% paraformaldehyde for 15 min. Then, 0.5% Triton™ X−100 was employed for 20 min to permeabilize the nuclear membrane, and used 5% normal serum for blocking at 25°C for 1 h. Finally, cells were stained with a Cell-Light™ EdU Apollo®488 in vitro Imaging Kit (Life Technologies, New York, USA). Staining was observed under a Pannoramic MIDI digital slide scanner (3DHISTECH, Ltd. Budapest, Hungary). The level of proliferation was represented as a percentage of the EdU-positive cells to the total cells.
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