Cell viability was monitored using Cell Proliferation Reagent Kit I (MTT; Roche Applied Science, Penzberg, Germany). The H1299 or H1975 cells transfected with si-AGAP2-AS1 (3000 cells/well) were grown in 96-well plates. Cell viability was assessed every 24 h following the manufacturer's protocol. All experiments were performed in quadruplicate. For each treatment group, wells were assessed in triplicate. For EdU assay, cells were cultured in 24-well plates, and 10 μM of EdU was added to each well. After 2 h, the cells were fixed with 4% formaldehyde for 30 min. After washing, EdU can be detected with a Click-iTR EdU Kit for 30 min, and the cells were stained with DAPI for 5 min and visualized using a fluorescent microscope (Olympus, Tokyo, Japan). The EdU incorporation rate was counted using Image-Pro Plus (IPP) 6.0 software (Media Cybernetics, Bethesda, MD, USA).
Click itr edu kit
Manufactured by Olympus
Sourced in Japan
The Click-iTR EdU Kit is a laboratory tool used to detect and measure cell proliferation. It utilizes a chemical reaction to incorporate a modified nucleoside, EdU, into newly synthesized DNA, allowing for the identification and quantification of proliferating cells.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescent microscope, by Olympus (7 mentions)
Image pro plus 6, by Media Cybernetics (3 mentions)
Mtt kit, by Merck Group (2 mentions)
Celltiter glor luminescent cell viability assay, by Promega (2 mentions)
Celltiter glo luminescent cell viability assay, by Promega (2 mentions)
Cell proliferation reagent kit 1, by Roche (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Celltiter glo, by Promega (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
9 protocols using click itr edu kit
1
Quantifying Cell Viability and Proliferation
2
Cell Proliferation and Colony Formation Assays
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Cell viability was tested with an MTT kit (Sigma) according to the manufacturer’s instructions. For the colony formation assay, a certain number of transfected cells were placed in each well of 6-well plates and maintained in proper media containing 10% FBS for 2 weeks, during which the medium was replaced every 4 days. Colonies were then fixed with methanol and stained with 0.1% crystal violet (Sigma) in PBS for 15 min. Colony formation was determined by counting the number of stained colonies. For the EdU incorporation assay, cells were cultured in 24-well plates; 10 μM EdU was added to each well, and cells were cultured for an additional 2 h. Then the cells were fixed with 4% formaldehyde for 30 min. After washing, EdU could be detected with a Click-iTR EdU Kit for 30 min, and the cells were stained with DAPI for 10 min and visualized using a fluorescent microscope (Olympus, Tokyo, Japan). The EdU incorporation rate was expressed as the ratio of EdU-positive cells to total DAPI-positive cells (blue cells), which were counted using Image-Pro Plus (IPP) 6.0 software (Media Cybernetics, Bethesda, MD, USA).
3
Cell Viability, Colony Formation, and EdU Assays
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Cell viability and colony-formation assay were performed as previously reported.41 (link) Briefly, cell viability was tested with MTT kit (Sigma) according to the manufacturer’s instruction. For colony-formation assay, a certain number of transfected cells were placed in each well of 6-well plates and maintained in proper media containing 10% FBS for two weeks, during which the medium was replaced every 3 days. Colonies were then fixed with methanol and stained with 0.1% crystal violet (Sigma) in PBS for 15 min. Colony formation was determined by counting the number of stained colonies. For the EdU incorporation assay, cells were cultured in 24-well plates. Then, 10 μM EdU was added to each well and the cells were cultured for an additional 2 hr. Then, the cells were fixed with 4% formaldehyde for 30 min. After washing, EdU can be detected with a Click-iTR EdU Kit for 30 min, and the cells were stained with DAPI for 10 min and visualized using a fluorescent microscope (Olympus). The EdU incorporation rate was expressed as the ratio of EdU-positive cells to total DAPI-positive cells (blue cells), which were counted using Image-Pro Plus (IPP) 6.0 software (Media Cybernetics).
4
Cell Proliferation Assay Protocol
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For cell counting kit‐8 (CCK‐8), after 48 h of transfection, a total of approximately 2 × 103 cells/well was seeded in 96‐well plate. After culturing at indicated time (0, 1, 2, 3, and 4 day), the cellular proliferation was detected using CellTiter‐GloR Luminescent Cell Viability Assay (Promega, Madison, WI, USA) according to manufacturer's instructions.
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well, and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click‐iTR EdU Kit, and images were visualized using fluorescent microscope (Olympus).
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well, and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click‐iTR EdU Kit, and images were visualized using fluorescent microscope (Olympus).
5
Cellular Proliferation Assays: CellTiter-Glo® and EdU
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CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, WI, U.S.A.), in accordance with the manufacturer's instructions, was used to measure cellular proliferation 48 hours after transfection for the Cell Counting Kit-8 (CCK-8) or ethynyl deoxyuridine (EdU) incorporation assay. The cells were fixed with 4 percent formaldehyde for 30 minutes after EdU (10 mM) was introduced to each well. Following washing, the Click-iTR EdU Kit was used to detect EdU, and fluorescence microscope images were used to view the results (Olympus). The following parameters were tested using kits and absorbance sing luminescent cell viability assay: CellTiter-Glo® (Promega, Madison, WI, U.S.A.)
6
TNBC Cell Viability Analyzed by EdU Assay
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TNBC cell viability was detected using an EdU assay. In brief, after transfection with the corresponding shRNA or plasmids, approximately 2 × 103 TNBC cells were incubated with 100 μl of 50 μM EdU per well for 2 h at 37 °C. After culturing, cell viability was assessed using the Click-iTR EdU Kit in accordance with the manufacturer’s instructions, and the cells were stained with DAPI for 10 min and visualized using a fluorescence microscope (Olympus). The viability was calculated based on the ratio of EdU-positive cells to the total DAPI-positive cells (blue cells).
7
Cell Proliferation Assay Using CCK-8 and EdU
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For cell counting kit-8, after 48 h of transfection, a total of approximately 2 × 10 3 cells/well was seeded into 96-well plate. After culturing at indicated time, the cellular proliferation was detected using CellTiter-GloR Luminescent Cell Viability Assay (Beyotime) according to manufacturer's instructions.
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click-iTR EdU Kit and images were visualized using fluorescent microscope (Olympus).
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click-iTR EdU Kit and images were visualized using fluorescent microscope (Olympus).
8
Cell Proliferation Analysis via Luminescent Assay
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After 48 h of transfection, a total of approximately 2 × 103 cells/well was seeded in 96-well plate. After culturing at indicated time (0, 6, 12, 24, and 48 day), the cellular proliferation was detected using CellTiter-Glo Luminescent Cell Viability Assay (Promega, Madison, WI, U.S.A.) according to manufacturer's instructions. EdU (10 mM) was added to each well, and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click-iTR EdU Kit, and images were visualized using fluorescent microscope (Olympus).
9
Cell Proliferation Assays with CCK-8 and EdU
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For Cell Counting Kit-8 (CCK-8), after 48 h of transfection, a total of approximately 2 × 10 3 cells/well was seeded in 96-well plate. After culturing at indicated time (0, 6, 12, 24 and 48 days), the cellular proliferation was detected using CellTiter-GloR Luminescent Cell Viability Assay (Promega, Madison, WI) according to manufacturer's instructions.
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click-iTR EdU Kit and images were visualised using fluorescent microscope (Olympus).
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click-iTR EdU Kit and images were visualised using fluorescent microscope (Olympus).
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