Cell proliferation was determined using Cell Counting Kit-8 (CCK-8) solution (Dojindo, Gaithersburg, MD, USA) and the cell-light 5-ethynyl-20-deoxyuridine (EdU) Apollo Imaging Kit (Ribobio, Guangzhou, China) in accordance with the manufacturer's protocol. For CCK-8 assay, cells were seeded at a concentration of 4 × 103 cells/well into three replicate wells on a 96-well plate and treated with 10 μL/well of CCK-8 solution during the last 4 h of culture daily for four consecutive days. The absorbance at 450 nm was measured with a microplate reader. The EdU assay was performed according to manufacturer's instructions. After EdU incubation, cells were treated with 1× Apollo solution and then stained with Hoechst 33342 (Ribobio, Guangzhou, China). The EdU positive cells was visualized under a fluorescence microscope (Olympus, Tokyo, Japan) and the positive percentage was defined as the proliferation rate. All reactions were performed in duplicate.
Edu apollo imaging kit
Manufactured by RiboBio
Sourced in China
The EdU (5-ethynyl-2'-deoxyuridine) Apollo Imaging Kit is a fluorescence-based cell proliferation assay that enables the detection and quantification of DNA synthesis in proliferating cells. The kit provides all the necessary components for the labeling, detection, and imaging of EdU-incorporated cells.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microscope, by Olympus (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Hoechst 33342, by RiboBio (1 mentions)
Cell counting kit 8 cck 8 solution, by Dojindo Laboratories (1 mentions)
Apollo solution, by RiboBio (1 mentions)
Ab27278, by Abcam (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Sc 365175, by Santa Cruz Biotechnology (1 mentions)
Ang 2, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Ab5694, by Abcam (1 mentions)
Dulbecco modified eagle medium (dmem), by ScienCell (1 mentions)
Cell light edu apollo in vitro imaging kit, by RiboBio (1 mentions)
Cell counting kit 8 cck 8 assay kit, by Dojindo Laboratories (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin and streptomycin, by Merck Group (1 mentions)
Cck 8 cell proliferation viability assay kit, by Merck Group (1 mentions)
Ambion paris system, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
9 protocols using edu apollo imaging kit
1
Cell Proliferation Assay Protocol
2
EdU Assay for Cell Proliferation
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Cell proliferation ability was measured by EdU assay [26 (link)]. The EdU Apollo Imaging Kit (RiboBio) was used in this study. Fifty μM/well EdU was added, and then GC cells were exposed to 4% paraformaldehyde for 0.5 h. Next, cells were incubated with Apollo solution (RiboBio) and diamidine phenylindole (DAPI; RiboBio). Cell pictures were saved by a microscopy (20 × objective lenses).
3
EdU Proliferation Imaging Protocol
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Cell-light 5-ethynyl-20-deoxyuridine (EdU) Apollo Imaging Kit (RiboBio, China) was used for measuring the cell proliferation according to manufacturer’s instructions. The percentage of EdU cells was assessed using a fluorescence microscope.
4
Sirtuin 3 Overexpression Regulates Angiogenesis
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Sirtuin 3 overexpressing lentivirus was purchased from GenePharma (GenePharma). Mini‐osmotic pump was purchased from Alzet (Durect). Ang II was purchased from Sigma (Sigma Aldrich). Mouse VEGFC ELISA kit was purchased from Meimian (Meimian). Lipofectamine 2000 was purchased from Invitrogen (Thermo Fisher). DMEM was purchased from Sciencell (Sciencell). Foetal bovine serum was purchased from Gibco (Thermo Fisher). EDU Apollo Imaging Kit was purchased from Ribobio (Ribobio). Antibodies against SIRT3, tubulin and GAPDH were purchased from Cell Signaling Technology (D22A3, 5335S and 5174S, CST). Antibody against ERK, p‐ERK and MYH6 was purchased from Proteintech (16443‐1‐AP, 80031‐1‐AP and 22281‐1‐AP, ProteinTech). Antibodies against LYVE1, VEGFR3 and α‐SMA were purchased from Abcam (ab19147, ab27278 and ab5694, Abcam). Mouse monoclonal antibody against SIRT3 was purchased from Santa Cruz (sc‐365175).
5
Quantifying Cancer Cell Proliferation
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Cell proliferation was determined using a Cell Counting Kit-8 (CCK-8) assay kit (Dojindo, Japan) and a cell-light 5-ethynyl-2′-deoxyuridine (EdU) Apollo Imaging Kit (Ribobio, Guangzhou, China). For the CCK-8 assay, U87 and U251 cells were seeded into 96-well plates for 0, 24, 48, and 72 h at a density of 3000 cells per well. Then, 10 μL CCK-8 solution was added to each well and incubated with the cells for 2 h. Absorbance was detected at 450 nm using a microplate reader (Bio-Rad, Hercules, CA, USA). EdU immunocytochemistry staining was performed by using a Cell-Light™ EdU Apollo In Vitro Imaging Kit (Ribobio, Guangzhou, China) at 24 h after the cell was plated into 96-well plates. The EdU-positive cells were visualized under a fluorescence microscope (Olympus, Tokyo, Japan).
6
Comprehensive lincRNA-p21 study protocol
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Ambion® PARIS™ system (PARIS™ Instruction Manual, Thermo Fisher Scientific, Waltham, MA, USA), BCA Protein Assay Kit (Thermo Fisher Scientific), CCK-8 Cell Proliferation/Viability Assay Kit (Sigma-Aldrich, St. Louis, MO, USA) and EdU Apollo Imaging Kit (RiboBio, Guangzhou, China), Cell Cycle (Nanjing KeyGEN, China), keyFluor 647-Annexin V/7-AAD Apoptosis Detection Kit (Nanjing KeyGEN, China), dimethyl sulfoxide (DMSO) (Sigma-Aldrich, fetal bovine serum (Gibco, Waltham, MA, USA), LincRNA-p21-lentivirus and negative control (Shandong ViGene Biosciences, Shandong, China), Matrigel (BD, Franklin Lake, NJ, USA), nuclear/cytosol fractionation kit (Biovision, San Francisco Bay, CA, USA), PBS (Gibco), PrimeScript RT Master Mix Kit (Takara, Shiga, Japan), penicillin and streptomycin (Sigma-Aldrich), RNase-Free DNase I and Purification Kit (Tiangen, Beijing, China), RiboTM lncRNA FISH Probe Mix and RiboTM Fluorescent In Situ Hybridization Kit (RiboBio, Guangzhou, China), RPMI1640 (Hyclone, Logan, UT, USA), SYBR Premix Ex Taq II Kit (Takara, Kusatsu, Japan), TRIzol reagent (Invitrogen, USA), trypsin and streptomycin (Nanjing Sunshine, Nanjing, China) were used. All primers used for this study, including LincRNA-p21, p21, β-actin, and U6, were purchased from Nanjing GenScript Company (Nanjing, China).
7
EdU Assay for Cell Proliferation
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An EdU Apollo Imaging kit (RiboBio, Guangzhou, Guangdong, China) was employed to conduct the EdU assays (31 (link)). Specifically, the miR-1224 mimics or negative control (NC) mimics-transfected SGC-25 or SGC-7901 cells (about 4000 cells per well) were planted into 48-well plates. Twenty-four hours later, EdU solution (50 μM final concentration) was added to each well. The treated cells were maintained for an additional 2 hours and added with DAPI solution (32 (link)). Finally, the cells were fixed with formaldehyde (4%) and observed using fluorescence microscopy.
8
Quantitative Cell Proliferation Assay
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The cell proliferation was examined using cell-light 5-ethynyl-20-deoxyuridine (EdU) Apollo Imaging Kit (RiboBio, China). A fluorescence microscopy was applied for the measurement of the rate of EdU-positive cells.
9
Quantifying Proliferating Cells with EdU
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Proliferating EC9706 cells were determined using the cell-light 5-ethynyl-20-deoxyuridine (EdU) Apollo Imaging Kit (RiboBio, China) according to the manufacturer’s protocol. EdU is a nucleoside analog of thymidine and is readily incorporated into DNA during active DNA synthesis only by proliferating cells (Salic and Mitchison, 2008 (link)). Cells were incubated with 50 μM EdU for 3 h after transfection. Subsequently, fixation, permeabilization, and EdU staining were performed. Nucleic acids in all cells were stained with Hoechst 33342, resulting in blue fluorescence. Proliferating cells were stained by conjugated reaction of Apollo dye and EdU, resulting in red fluorescence. All images were obtained by a fluorescence microscope (FSX100, Olympus, Japan).
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