After transfection, GC cells were seeded onto 96-well plates. The EdU (5-ethynyl-20-deoxyuridine) Apollo-567 Kit (RIBOBIO, Shanghai, China) was used to quantify the proliferation ability of indicated GC cells. EdU and DAPI dyes were exploited to treat GC cells. A fluorescent microscope was used for cell visualization (Olympus, Tokyo, Japan). The experiment was repeated for three times.
Apollo 567 kit
Manufactured by RiboBio
Sourced in China
The Apollo-567 Kit is a laboratory equipment product designed for DNA and RNA analysis. The kit includes essential components for performing various nucleic acid manipulation and detection procedures. The core function of the Apollo-567 Kit is to facilitate the processing, amplification, and analysis of genetic material samples.
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12 protocols using apollo 567 kit
1
Quantifying GC Cell Proliferation
2
Proliferation Assay for NSCLC Cells
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NSCLC cells were seeded into six-well plates at 8×104 cells per well, and cultured for 48h. An EdU (5-ethynyl-20-deoxyuridine) Apollo-567 Kit (RiboBio, Guangzhou, China) was applied. EdU and DAPI dyes were added to NSCLC cells for additional 4h. Under a fluorescent microscope cell nuclei were visualized. For each condition total 800 nuclei in five random views were included to calculate the EdU ratio (EdU/DAPI×100%). For cell counting assay 1×104 NSCLC cells (with applied genetic treatments) per well were initially seeded (at 0h), cells were cultured for applied time periods with cell number recorded.
3
EdU Proliferation Assay in OS Cells
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OS cells were seeded into six-well plates at 1.2×105 cells per well and were cultured for 48 h. An EdU (5-ethynyl-20-deoxyuridine) Apollo-567 Kit (RiboBio, Guangzhou, China) was applied to quantify cell proliferation according to the attached protocols. EdU and DAPI fluorescent dyes were added to OS cells and the cells were visualized under a fluorescent microscope (Leica, Shanghai, China). Five random views (n = 5) with a total of 1 000 cells per each condition were used to calculate the average EdU ratio (% vs. DAPI).
4
Evaluating Cell Growth and Proliferation with CCK-8 and EdU
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Cell counting kit-8 (CCK-8) (Vazyme Biotech, Nanjing, China) is applied to evaluate the growth of cells. In brief, 2 × 103 cells are seeded into cell cultural plate and activated by LPS (1 μg/ml) for 12, 24, and 48 hrs. The absorption is determined following the protocol of CCK-8. CellLight 5-ethynyl-2′-deoxyuridine (EdU) is also carried out by use of Apollo567 kit (RiboBio, Guangzhou, China) for cell proliferation determination after stimulation by LPS for 24 hrs.
5
Cell Proliferation Quantification using EdU
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Cells were seeded into six-well plates at 1 × 105 cells per well. Following the applied treatments, an EdU (5-ethynyl-20-deoxyuridine) Apollo-567 Kit (RiboBio, Guangzhou, China) was utilized to quantify cell proliferation. EdU and DAPI were both added to the cultured cells, and visualized under a fluorescent microscope. EdU ratio (% vs. DAPI) was calculated.
6
Quantifying U2OS Cell Proliferation
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U2OS cells were initially seeded into six-well plates (at 8 × 104 cells per well), following the treatment applications and cultured for 48 h; an EdU (5-ethynyl-20-deoxyuridine) Apollo-567 Kit (RiboBio, Guangzhou, China) was utilized to quantify cell proliferation. EdU and DAPI dyes were added to the U2OS cells and visualized under a fluorescent microscope. Nuclear EdU ratio was calculated from 800 nuclei in five random views (1 × 200) in each condition.
7
Cell Proliferation Assay with EdU
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EdU (5-ethynyl-20-deoxyuridine) Apollo-567 Kit (RIBOBIO, Shanghai, China) was employed to test cell proliferation. Briefly, cells were seeded onto the twelve-well plates. Following treatment, EdU and DAPI dyes were added, and cells were incubated for additional 8h. Cell nuclei were then visualized under a fluorescent microscope. For each condition at least 500 cells in five random views were included to calculate EdU ratio (EdU/DAPI×100%).
8
Glioma Cell Proliferation Assay
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Cells were seeded into six-well plates at 6 × 104 cells per well, and cultured for 48 h. An EdU (5-ethynyl-20-deoxyuridine) Apollo-567 Kit (RiboBio, Guangzhou, China) was applied. EdU and DAPI dyes were added to glioma cells for additional 4 h. Under a fluorescent microscope cell nuclei were visualized. For each condition total 300 nuclei in five random views were included to calculate EdU ratios (EdU/DAPI × 100%).
9
Quantifying Cell Proliferation with EdU Assay
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A Cell-Light EdU (5-ethynyl-2′-deoxyuridine) Apollo 567 Kit (Ribobio, Guangzhou, China) was carried out to quantify cell proliferation. Briefly, NSCLC cells were seeded into 96-well plates at 5×103 cells/well. After 48 h, 100 μl medium containing 10 μM EdU was added into each well. Cells were incubated for 2 h, fixed, and co-stained with DAPI. EdU, and DAPI staining was captured by a fluorescence microscopy (Nikon, Japan). For each treatment, five random views with a total of 1,000 nuclei were included to calculate the average EdU ratio (% vs. DAPI).
10
NSCLC Cell Proliferation Assay
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NSCLC cells (at 1×105 cells per well) were seeded into six-well plates and cultured for 72h. An 5-ethynyl-20-deoxyuridine(EdU) Apollo-567 kit (RiboBio, Guangzhou, China) was utilized to test and quantify cell proliferation, with EdU ratio (EdU/DAPI×100%) determined from at least 500 nuclei in five random views (1×100) of each condition.
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