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Edu apollo dna in vitro kit

Manufactured by RiboBio
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Sourced in China

The EdU Apollo DNA in vitro kit is a laboratory tool used for the detection and quantification of DNA synthesis in cell-free systems. The kit utilizes a chemical compound called EdU (5-ethynyl-2'-deoxyuridine) to label newly synthesized DNA, which can then be visualized and analyzed using the provided reagents and protocols.

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Lab products found in correlation

Microplate reader, by Bio-Rad (4 mentions) Beyoclick edu cell proliferation kit with dab, by Beyotime (3 mentions) Automated cell counter, by Countstar (3 mentions) Glass bottom cell culture dish, by NEST Biotechnology (2 mentions) Fluorescence microscopy, by Olympus (2 mentions) Cell counting kit 8 (cck8), by Transgene (2 mentions) Beyoclick edu cell proliferation kit, by Beyotime (1 mentions) Crystal violet, by Solarbio (1 mentions) Cell counter, by Countstar (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Multiskan mk3, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Beyotime (1 mentions) Crystal violet, by Beyotime (1 mentions) E0226, by Beyotime (1 mentions) Cck 8 assay, by Beyotime (1 mentions) Annexin 5 fitc pi double staining kit, by Dojindo Laboratories (1 mentions) Cell counting kit 8 (cck8), by Boster Bio (1 mentions) Hemocytometer, by Corning (1 mentions) Fv10i, by Olympus (1 mentions) β glycerophosphate, by Merck Group (1 mentions)

Close spelling variants of this product

EdU kits (10 citations) Cell-Light Edu Apollo DNA in vitro Kit (3 citations) Apollo DNA in vitro kit (2 citations) EdU Apollo-DNA kit (2 citations)

55 protocols using edu apollo dna in vitro kit

1

EdU Assay for Cell Proliferation

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EdU Apollo DNA in vitro kit (Ribobio, Guangzhou, China) was used to detect cell proliferation. Cells were plated into a glass-bottom cell culture dish (NEST, Hong Kong, China) at a density of 2.0 × 105. Briefly, cells were fixed with 4% paraformaldehyde (m/v) for 30 min, and followed by incubation of 30 μM EdU at 37 °C for 90 min. After permeabilized in 0.5% Triton X-100, the Apollo staining solution was added into the cell culture medium for 30 min in the dark. Finally, the cells were incubated with 20 μg/mL 4′,6-diamidino-2-phenylindole (DAPI) for 10 min. The EdU index (%) was the average ratio of the number of EdU-positive cells over total cells in five randomly selected areas under the confocal laser scanning microscope (FV10i).
Yuan Y., Yan G., He M., Lei H., Li L., Wang Y., He X., Li G., Wang Q., Gao Y., Qu Z., Mei Z., Shen Z., Pu J., Wang A., Zhao W., Jiang H., Du W, & Yang L. (2021). ALKBH5 suppresses tumor progression via an m6A-dependent epigenetic silencing of pre-miR-181b-1/YAP signaling axis in osteosarcoma. Cell Death & Disease, 12(1), 60.
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2

Cell Proliferation Assay with EdU

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Cell proliferation was determined by the ethynyl-2-deoxyuridine incorporation assay using an EdU Apollo DNA in vitro kit (RIBOBIO, Guangzhou, China) according to the manufacturer’s instructions. Briefly, after transfection with the corresponding vector, cells were incubated for 2 h at 37 °C, with 100 µL of 50 µM EdU/well. The cells were identified using fluorescence microscopy. Each experiment was carried out three times.
Zhang R.N., Wu D.M., Wu L.P, & Gao G.W. (2021). LncRNA LINC00337 sponges mir-1285-3p to promote proliferation and metastasis of lung adenocarcinoma cells by upregulating YTHDF1. Cancer Cell International, 21, 550.
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3

Detecting Cellular DNA Synthesis via EdU

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Detection of EdU incorporation into the DNA was performed with an EdU Apollo DNA in vitro kit (Guangzhou RiboBio Co., Ltd.). According to the manufacturer's instructions, 50 µM EdU was used for incorporation. The established cells were seeded in 96-well plates and incubated overnight at 37°C. The supernatant was removed by aspiration, and the attached cells were fixed with 100 µl fixing buffer (4% polyformaldehyde in PBS) for 30 min at room temperature. Following incubation with 2 mg/ml glycine for 10 min at room temperature, the cells were washed with 1X PBS. The cells were treated with 100 µl/well permeabilization buffer (1X PBS containing 0.5% Triton X-100) for 10 min at room temperature and incubated with 100 µl 1X Apollo solution for 30 min at room temperature in the dark. Subsequently, cells were incubated with 100 µl 1X Hoechst 33342 solution for 30 min at room temperature in the dark. Finally, the cells were observed under a fluorescence microscope.
Chen R., Sheng C., Ma R., Zhang L., Yang L, & Chen Y. (2021). PLAC1 is an independent predictor of poor survival, and promotes cell proliferation and invasion in cervical cancer. Molecular Medicine Reports, 24(5), 800.
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4

Quantitative Analysis of Cell Proliferation

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EdU assays were performed using an EdU Apollo DNA in vitro kit (Ribobio, China), according to the manufacturer’s instructions. Briefly, the transfected cells were plated in 24-well plates overnight. 30 μM EdU were added into each well at 37°C for 90 min. After fixation with 4% paraformaldehyde and incubation in 0.5% Triton X-100, the cells were stained using Apollo for 30 min.
Lastly, the cells were incubated with DAPI and observed under a fluorescence microscope. The number of EdU-positive cells was measured using image J.
Sun Z., Sun X., Qin G., Li Y., Zhou G, & Jiang X. (2023). FTO promotes proliferation and migration of bladder cancer via enhancing stability of STAT3 mRNA in an m6A-dependent manner. Epigenetics, 18(1), 2242688.
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5

Proliferation Assay of Human HCC Cells

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An EdU Apollo DNA in vitro kit (RIBOBIO, Guangzhou, China) was used to determine the cell proliferation. The entire operation process followed the manufacturer’s instructions. Exponentially growing human HepG2 and Huh-7 cells were placed on 13 mm glass coverslips, respectively. Then, CDCA4-siRNA and the negative control (NC) were transfected into human HCC cells with LipofectamineTM2000, respectively. After cultured 24 h, the cells were added to 50 μM EdU labeling media and then incubated for 2 hours at RT under 5% CO2. Subsequently, cells were treated with 4% paraformaldehyde, glycine, 1% Triton X-100. Apollo and Hoechst staining were performed for 30 min respectively. The images were taken by fluorescence microscopy (Olympus, Tokyo, Japan).
Fang H., Sheng S., Chen B., Wang J., Mao D., Han Y., Liu Y., Wang X., Gui S., Zhang T., Zhang L., Li C., Hu X., Deng W., Liu X., Xu H., Xu W., Wang X., Liu R, & Kong W. (2022). A Pan-Cancer Analysis of the Oncogenic Role of Cell Division Cycle-Associated Protein 4 (CDCA4) in Human Tumors. Frontiers in Immunology, 13, 826337.
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6

EdU Proliferation Assay Protocol

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Cell proliferation was also evaluated by EdU incorporation assay using an EdU Apollo DNA in vitro kit (RiboBio, Guangzhou, China). Briefly, cells after transfection were incubated in 96-well plates (5,000 cells per well) for 24 h at 37°C and next subjected with 100 μL of 50 μM EdU per well. After culturing cells with EdU for 12 h, cells were fixed and then counterstained with DAPI. The EdU staining was observed via a fluorescence microscopy (Mshot, Guangdong, China).
Feng L., Liu T., Shi J., Wang Y., Yang Y., Xiao W, & Bai Y. (2023). Circ-UBR4 regulates the proliferation, migration, inflammation, and apoptosis in ox-LDL-induced vascular smooth muscle cells via miR-515-5p/IGF2 axis. Open Medicine, 18(1), 20230751.
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7

Cell Proliferation Quantification via EdU Assay

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Cell proliferation was determined by a EdU Apollo DNA in vitro kit (Ribobio, China). Briefly, after fixing with 4% paraformaldehyde (m/v) for 30 min, the cells (2 × 105/mL seeded in a 24-well plate) were treated with 30 μM/mL EdU at 37 °C for 90 min. After permeabilization in 0.5% Triton X-100, the Apollo staining solution was added to the plate in dark for 30 min. Then, the cells were incubated with 4′,6-diamidino-2-phenylindole (DAPI, 20 μg/mL) for 10 min. The average ratio of EdU-positive cells to total cells was calculated in randomly selected areas under a microscope (Olympus).
Yuan Y., Mei Z., Qu Z., Li G., Yu S., Liu Y., Liu K., Shen Z., Pu J., Wang Y., Wang C., Sun Z., Liu Q., Pang X., Wang A., Ren Z., Wang T., Liu Y., Hong J., Xie J., Li X., Wang Z., Du W, & Yang B. (2023). Exosomes secreted from cardiomyocytes suppress the sensitivity of tumor ferroptosis in ischemic heart failure. Signal Transduction and Targeted Therapy, 8, 121.
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8

Cell Proliferation Assay using EdU

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Cell proliferation ability was also tested by EdU Apollo DNA in vitro kit (RIBOBIO, China) according to instructions. MGC-803 and MKN-45 were seeded in a 96-well plate 24h before the transfection of either si-lincRNA-p21 or si-NC. At the 48h time point, 100 μl of 50 μM EdU was added in each well and incubated for 2 h at 37%. After that, the cells were fixed in 100ul of 4% paraformaldehyde (prepared in PBS) for 30 minutes at room temperature. The cells were incubated in 50 μl of 2 mg/ml glycine for 5 minutes followed by PBS washing. Afterwards, the cells were permeabilized with 1% TritonX and reacted with 1X Apollo solution for 30 minutes at the room temperature in dark subsequently. Finally, 40μl of Hoechst solution was added in each well and incubated for 15 minutes at room temperature in dark with PBS washing followed. In the end, the cells were visualized and photographed under a fluorescence microscopy. Experiments were repeated at least three times.
Chen Y., Wei G., Xia H., Yu H., Tang Q, & Bi F. (2017). Down regulation of lincRNA-p21 contributes to gastric cancer development through Hippo-independent activation of YAP. Oncotarget, 8(38), 63813-63824.
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9

Cell Proliferation and Colony Formation

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Cell proliferation was determined by an ethynyl-2-deoxyuridine (EdU) incorporation assay using an EdU Apollo DNA in vitro kit (RiboBio, Guangzhou, China) and BeyoClick™ EdU Cell Proliferation Kit with DAB (Beyotime, China). For cell clone formation experiment, 200 OSRC2 and Caki-1 cells were seeded in 6-well plates. After culturing for 2 to 3 weeks, stain with 0.5% crystal violet for 10 min and count the number of clones under a microscope. Experiments were repeated at least three times.
Yang W., Zhang K., Zhang Z., Zhou J., Li L., Xu Y., Qiu J., Cai L., Gong Y, & Gong K. (2022). Claudin-10 overexpression suppresses human clear cell renal cell carcinoma growth and metastasis by regulating ATP5O and causing mitochondrial dysfunction. International Journal of Biological Sciences, 18(6), 2329-2344.
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10

Quantifying Cell Proliferation with EdU Assay

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Cell proliferation was determined by an ethynyl-2-deoxyuridine incorporation assay using an EdU Apollo DNA in vitro kit (RIBOBIO, Guangzhou, China) following the manufacturer’s instructions. Briefly, 5 × 104 cells/well were seeded in a 24-well plate for 24 h, and then the cells were incubated with 100 μl of 50 μM EdU per well for 2 h at 37°C. Then, the cells were fixed for 30 min at room temperature using 100 μl of a fixative buffer (4% polyformaldehydein PBS). Subsequently, the cells were incubated with 50 μl of 2 mg/mL glycine for 5 min, followed by washing with 100 μl of PBS. After permeabilization with 0.5% Triton X-100, the cells were reacted with a 1X Apollo solution for 30 min at room temperature in the dark. After that, the cells were incubated with 100 μl of a 1X Hoechst 33,342 solution for 30 min at room temperature in the dark, followed by washing with 100 μl of PBS. The cells were then visualized by fluorescence microscopy. Experiments were repeated at least three times.
Bao Z., Zhan Y., He S., Li Y., Guan B., He Q., Yang X., Li X., Fang D, & Zhou L. (2019). Increased Expression Of SOX2 Predicts A Poor Prognosis And Promotes Malignant Phenotypes In Upper Tract Urothelial Carcinoma. Cancer Management and Research, 11, 9095-9106.
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