We investigated the proliferation of OS cells and OS cells treated with conditioned medium or indirect co-culture for 4 days using a Cell-Light 5-Ethynyl-2′- deoxyuridine (EdU) [46 (link)] Cell Proliferation Kit (Guangzhou RiboBio, Guangzhou, China), according to the manufacturer's instructions. Briefly, OS cells were seeded in a 96-well plate, and the medium was replaced with 100 μL of 10 μM EdU medium in each well. The cells were then incubated for 2 h and fixed with 4% paraformaldehyde. The formaldehyde was neutralized with 2 mg/mL glycine solution, after which the cells were subjected to 0.5% Triton X-100 permeabilization. Then, the cells were stained with Apollo® 567 and incubated for 30 min before being permeabilized with 0.5% Triton X-100. The cells were subsequently counterstained with Hoechst 33342 and imaged via fluorescence microscopy. EdU-labelled cells in each well were counted manually in ten randomly selected fields of view, and the percentages of EdU-positive cells were used to determine cell proliferative activity. All assays were performed in triplicate.
Cell light 5
Manufactured by RiboBio
Sourced in China
Cell-Light 5- is a laboratory equipment designed for cell culture applications. It is a multi-functional instrument that facilitates various cell-based experiments and analyses. The core function of Cell-Light 5- is to provide a controlled environment for the cultivation and observation of cells.
Automatically generated - may contain errors
Lab products found in correlation
Apollo567 in vitro kit, by RiboBio (4 mentions)
Edu dna cell proliferation kit, by RiboBio (1 mentions)
3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2h tetrazolium bromide mtt, by Merck Group (1 mentions)
Cck 8 detection kit, by Beyotime (1 mentions)
Ix73 aizfl ph, by Olympus (1 mentions)
Fluorescence microscope, by Leica camera (1 mentions)
Tunel assay, by Beyotime (1 mentions)
Annexin 5 pe 7 aad apoptosis detection kit, by BD (1 mentions)
Apollo 488 in vitro kit, by RiboBio (1 mentions)
Hoechst 33342, by Abcam (1 mentions)
10 protocols using cell light 5
1
Evaluating OS Cell Proliferation
2
EdU DNA Cell Proliferation Assay
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A Cell-Light 5-ethynyl-2-deoxyuridine (EdU) DNA Cell Proliferation Kit (RiboBio, Guangzhou, China) was utilized to determine cell viability. Briefly, treated LoVo cells were seeded in 24-well plates at a density of 1×105 cells per well, and then exposed to 30 µM EdU at 37 °C for 2 hrs. Next, the cells were fixed with 4% formaldehyde (cat. no. 153814; Biosharp, Wuhan, China) at room temperature for 30 mins, and then permeabilized by exposure to 0.5% Triton X-100 for 10 mins; after which, they were washed and stained with DAPI. After staining, cell fluorescence was measured by flow cytometry.
3
Measuring Cell Proliferation and Viability
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The Cell-Light 5-ethynyl-2-deoxyuridine (EdU) Apollo 567 Kits (RiboBio, Guangzhou, China) were used to detect cell proliferation. Cell proliferation rate = (EdU-positive cells)/(total cells) × 100% [54 (link)]. The 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) (Sigma, St. Louis, MO, USA) and CCK-8 detection kits (Beyotime, Shanghai, China) were adopted to determine cell viability [55 (link)–57 (link)].
4
Quantifying Cell Proliferation with EdU
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Cell growth was detected by Cell-Light 5-ethynyl-2′-deoxyuridine (EdU) Apollo 567 In vitro kit (RiboBio, Guangzhou, China). After transfection, the cells were cultured in a 96-well plate (1 × 104/well) for 48 h, followed by incubation with an EdU-labeled medium of 50 μM for 2 h. Then, the cells were immobilized in 4% paraformaldehyde, permeated in 0.5% TritonX-100/PBS, and stained using Apollo solution and Hoechst33342. An inverted fluorescence microscope IX73-AIZFL/PH (OLYMPUS Corporation, Japan) was used to generate cell images.
5
Comprehensive Analysis of Cell Proliferation and Apoptosis
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Cell proliferation was analyzed using a Cell-Light 5-ethynyl-2′-deoxyuridine (EdU) Apollo567 In Vitro Kit (RiboBio, Shanghai, China) in strict accordance with the manufacturer's instructions. Images were acquired at 20 × magnification with a fluorescence microscope (Leica, Solms, Germany). EdU-positive cells were counted in the selected cells and calculated as the mean value from multiple fields of view. Cell apoptosis assays were performed using an Annexin V-PE 7-AAD Apoptosis Detection Kit (BD Biosciences, CA, USA) and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL assay, Beyotime, Shanghai, China) according to the manufacturer's protocol. For Annexin V-PE 7-AAD Apoptosis assay, the cells were analyzed the percentage of apoptotic cells, that cells undergoing early and late apoptosis, among 10,000–20,000 counted cells using a flow cytometer (BD Biosciences, CA, USA). For TUNEL assay, images of apoptosis cells were obtained using a microscope (Leica, Solms, Germany).
6
EdU-based Cell Proliferation Assay
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Cell proliferation was analysed using Cell‐Light 5‐ethynyl‐2’‐deoxyuridine (EdU) Apollo 567 in vitro kit (Guangzhou RiboBio Co., Ltd.). The transfected cells were cultured in a 96‐well plate (1 × 104 cells/well). After 48 hours of culture, 50 μmol L−1 of EdU‐labelled medium was added, followed by incubation for an additional 2 hours. The cells were then fixed in 4% paraformaldehyde (PFA), permeabilized in 0.5% Triton×‐100/PBS, and stained with Apollo staining solution and Hoechst33342. Cell images were obtained under an inverted fluorescence microscope IX73‐AIZFL/PH (Olympus Optical Co., Ltd.).
7
EdU Incorporation Assay for Cell Proliferation
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EdU assay were performed using Cell-Light 5-ethynyl-2’-deoxyuridine (EdU) Apollo488 in vitro Kit (Ribobio, Guangzhou, China). Briefly, GC cells were plated in 6-well plates, and incubated with 1× EdU assay buffer for 2 h. After washing, the cells were incubated with Apollo®488 staining for 30 min. The EdU(+) cells were analyzed by flow cytometry.
8
Proliferative Ability of HUVECs
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The proliferative ability of HUVECs after different transfections or different exosomes treated was determined by Cell‐Light 5‐ethynyl‐2′‐deoxyuridine (EdU) Apollo in vitro image kit (RiboBio). After pretreatment as described above, HUVECs were incubated in 50 μM EdU for 2 h, and were then fixed and stained following the appropriate manufacturer's instructions.
9
Cell Proliferation Measurement via EdU Assay
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At 24 hours after transduction, cell proliferation was measured using a Cell-Light 5-ethynyl-2?-deoxyuridine (EdU) Apollo567 In Vitro Kit (Ribobio) according to the manufacturer's protocol, as previously described (Zheng et al., 2020). Briefly, the cells were exposed to EdU for 2 hours, then washed and fixed with 4% paraformaldehyde for 30 minutes. Following washing with phosphate-buffered saline, the cells were blocked with phosphate-buffered saline containing 0.5% Triton X-100 for 10 minutes. Cells were incubated with 200 μL of 1× Apollo® staining reaction solution for 30 minutes, and the nuclei were stained with Hoechst 33342 (Abcam, Cambridge, MA, USA, Cat# ab228551, 1:1000).
10
Cell Proliferation Assessment via EdU Labeling
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A commercial Cell-Light 5-ethynyl-2′-deoxyuridine (EdU) In Vitro Imaging Kit (RiboBio Co., C10310) was used to assess cell proliferation, according to the manufacturer's instructions. In brief, EdU was added to the cell culture medium (as indicated earlier) at a final concentration of 100 nmol/L for labeling and standardized permeabilization and fixation was done before incubating the cells with the reaction cocktail (30 min). After fluorescence microscopy, results were expressed as the percentage of EdU-positive cells in 6 independent experiments.
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