Yefluor 488 edu imaging kit

Manufactured by Yeasen

Sourced in China

The Yefluor 488 EdU Imaging Kit is a laboratory product designed for cell proliferation analysis. It contains reagents to detect and visualize cells undergoing DNA synthesis, a process associated with cell division. The kit utilizes the EdU (5-ethynyl-2'-deoxyuridine) labeling method, which allows for the detection of newly synthesized DNA. The Yefluor 488 dye is used to fluorescently label the incorporated EdU, enabling the visualization and quantification of proliferating cells.

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5 protocols using yefluor 488 edu imaging kit

ESCC Cell Proliferation Assays

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The cell counting kit‐8 (Dojindo,) was utilized to evaluate the proliferation of ESCC cells. The transfected cells were inoculated in 96‐well plates, and then 10 μl of the CCK8 solution was added at 24, 48 and 72 h, respectively, and the cells continued to be cultured for 4 h before the absorbance was detected at 450 nm.
5‐ethynyl‐2′‐deoxyuridine (EdU) assay was also employed in the assessment of ESCC cell proliferation. In this study, a Yefluor 488 EdU Imaging Kit (Yeasen) was used for analysis. ESCC cells were cultured in a 96‐well plate for 24 h. Then, the cells were incubated with EdU solution for 2 h. After the cells were fixed and stained with the nuclear dye DAPI for 30 min in the dark, the proliferation capacity was then observed and analyzed under the microscope.

Guan X., Guan X., Wang Y., Lan T., Cheng T., Cui Y, & Xu H. (2022). Circ_0003340 downregulation mitigates esophageal squamous cell carcinoma progression by targeting miR‐940/PRKAA1 axis. Thoracic Cancer, 13(8), 1164-1175.

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Cell Proliferation Imaging with EdU

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Cell proliferation was examined using Yefluor 488 EdU Imaging Kit (Yeasen, Shanghai, China). Cells were first incubated with EdU at 37 °C for 4 h. After fixation and permeabilization, the cells were reacted with 1 mL Click-iT reaction cocktail in darkness for 30 min. For nuclear staining, cells were incubated with DAPI. Finally, the fluorescence was observed under a fluorescence microscope (Nikon, Tokyo, Japan).

Miao Y., Zhang W., Liu S., Leng X., Hu C, & Sun H. (2021). HOXC10 promotes growth and migration of melanoma by regulating Slug to activate the YAP/TAZ signaling pathway. Discover. Oncology, 12, 12.

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Evaluating Breast Cancer Cell Proliferation

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Cell proliferation of breast cancer cells was assessed using Yefluor 488 EdU Imaging Kit (YEASEN, Shanghai, China) according to the manufacturer’s protocol. Cells were seeded into 24-well plate and incubated with 50 μM of EdU at 37 °C. After 4 h of incubation, cells were fixed with 4% paraformaldehyde solution at room temperature for 15 min. The cells were washed with PBS, and then permeabilized with 0.5% Triton X-100 at room temperature for 20 min. After washing with PBS, the cells were reacted with 1 mL of 1 mL Click-iT reaction cocktail in darkness for 30 min. Subsequently, cells were stained with 1 μg/mL of DAPI for 30 min and visualized under a fluorescence microscope (Nikon, Tokyo, Japan).

Wang Q., Liang D., Shen P., Yu Y., Yan Y, & You W. (2021). Hsa_circ_0092276 promotes doxorubicin resistance in breast cancer cells by regulating autophagy via miR-348/ATG7 axis. Translational Oncology, 14(8), 101045.

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Metabolic Profiling of TGF-β1-Induced EMT

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Tan IIA (purity > 98%), [U-¹³C]-glucose, l-serine, l-glycine, l-lactic acid, l-alanine, pyruvate, l-malic acid, succinate, fumarate, l-glutamic acid, citric acid, α-ketoglutaric acid, l-asparatic acid, and l-proline were purchased from Shanghai Yuanye Biotechnology Co., Ltd (Shanghai, China). Recombinant human TGF-β1, Yefluor 488 EdU imaging kit, and cell cycle and apoptosis analysis kit were purchased from Yeasen (Shanghai, China). Sulforaphane (SFN), dimethyl sulfoxide (DMSO) and methoxyamine hydrochloride were purchased from Sigma-Aldrich (St. Louis, MO, USA). d-glucose was purchased from Aladdin (Shanghai, China). Dulbecco's modified Eagle medium (DMEM), minimum essential medium (MEM), and 0.25% trypsin were purchased from Gibco (Carlsbad, CA, USA). Cell counting kit-8 (CCK-8) was purchased from Vazyme (Nanjing, China). SB 204990 was purchased from MedChemExpress (Princeton, NJ, USA). BTA was purchased from Macklin (Shanghai, China). Glucose assay kit was purchased from Elabscience (Wuhan, China). Lactate assay kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide was purchased from Acmec (Shanghai, China). Triton ×100, hoechst33342, antifade mounting medium and 4',6-diamidino-2-phenylindole (DAPI) were purchased from Beyotime (Shanghai, China).

Shan B., Zhou H., Guo C., Liu X., Wu M., Zhai R, & Chen J. (2023). Tanshinone IIA ameliorates energy metabolism dysfunction of pulmonary fibrosis using 13C metabolic flux analysis. Journal of Pharmaceutical Analysis, 14(2), 244-258.

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Cell Proliferation Quantification with EdU Assay

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5-Ethynyl-2ʹ deoxyuridine (EdU) assay was performed using the Yefluor 488 EdU Imaging Kit (Yeasen, China) according to the manufacturer's instructions. Briefly, PANC-1, MIA PaCa-2, and AsPC-1 cells were cultured in an imaging-appropriate dish and treated with DMSO and ST. Thereafter, cells were incubated with 50 μM EdU reaction reagent for 2 hours, washed with phosphate-buffered saline (PBS) two times, immobilized with 4% paraformaldehyde for 30 min, and incubated with Apollo staining reaction solution. Nuclei were stained with 4,6-diamino-2-phenylindole and visualized using a fluorescence microscope at 488 nm (magnification 400×).

Wang J., Zeng Z., Lei S., Han J., Liao S., Zhang J., Wang L., Dong Y., Li H, & Chen T. (2022). Sinapine Thiocyanate Inhibits the Proliferation and Mobility of Pancreatic Cancer Cells by Up-Regulating GADD45A. Journal of Cancer, 13(4), 1229-1240.

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