Apoptosis, cell cycle detection and MTT assay were performed as described previously 29 , 30 using the Multicaspase Kit (Millipore Corporation, Hayward, US), Cell Cycle Detection Kit (BD Pharmingen, San Diego, US) and Thiazolyl Blue Tetrazolium Bromide (MTT, Sigma‐Aldrich Corp.).
Multicaspase kit
Manufactured by Merck Group
Sourced in United States
The Multicaspase Kit is a laboratory instrument used for the detection and measurement of multiple caspase enzymes in cell samples. It provides a quantitative assessment of caspase activity, which is crucial for understanding cellular processes related to apoptosis and other forms of programmed cell death.
Automatically generated - may contain errors
Lab products found in correlation
7 aminoactinomycin d 7 aad, by Merck Group (2 mentions)
Muse cell analyzer, by Merck Group (2 mentions)
Cell cycle detection kit, by BD (1 mentions)
Pi rnase staining buffer, by BD (1 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions)
Flow cytometry, by Merck Group (1 mentions)
Cell cycle kit, by Merck Group (1 mentions)
6 protocols using multicaspase kit
1
Apoptosis, Cell Cycle, and MTT Assay
2
Apoptosis and Cell Cycle Analysis Protocol
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For cell apoptosis LECs (2×105) were seeded in 6-well plates, after being treated with different doses of THZ1 for 24h, cells were trypsinized and washed twice by PBS and then incubated with MultiCaspase kit (Millipore) for 30 min at 37°C in dark, then 7-aminoactinomycin D (7-AAD, Millipore) were added and incubated for 5 minutes in room temperature at dark 25 (link).
For cell cycle analysis, cells were fixed in cold 70% ethanol at 4°C overnight before incubated with staining solution (BD) at room temperature for 15 minutes following the manufacturer's instructions. After that, the fluorescence intensity was measured by MuseTM Cell Analyzer (Millipore).
For cell cycle analysis, cells were fixed in cold 70% ethanol at 4°C overnight before incubated with staining solution (BD) at room temperature for 15 minutes following the manufacturer's instructions. After that, the fluorescence intensity was measured by MuseTM Cell Analyzer (Millipore).
3
Cell Viability and Apoptosis Assay
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The Cell Counting Kit-8 (CCK8, Dojindo, Japan) was utilized to determine cell viability. Apoptosis of C2C12 cells, as assessed in the presence or absence of lidocaine, was performed with the use of the Multi-caspase Kit (Millipore, Cat: MCH100109, USA). PI/RNase Staining Buffer (BD Biosciences Pharmingen, Cat:550825, USA) was utilized in the cell cycle assay.
4
Curcumin Induces Apoptosis and Cell Cycle Alterations
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LECs treated with different concentrations of curcumin for 48 h were collected and washed with PBS; cell apoptosis and cell cycle were analyzed by flow cytometry (Millipore, Darmstadt, Hesse, Germany) with the MultiCaspase Kit and the Cell Cycle Kit (Millipore, Darmstadt, Hesse, Germany). All procedures were proceeded according to the manufacturer's protocols as we previously did [21 (link), 22 (link)]. The statistical analysis was performed with the software version 1.3 of the Muse™ Cell Analyzer (Millipore, Darmstadt, Hesse, Germany).
5
Quantifying Oxidative Stress and Apoptosis in RPE Cells
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ROS was examined using dihydroethidium (DHE; Millipore, EMD Millipore Corporation, Hayward, CA), according to the manufacturer's instructions. Briefly, RPE cells were seeded into 60 mm culture plates and split using 0.25% trypsin and washed with PBS. After gathering cells, DHE was added to each dish for 30 min at 37°C in dark. And then, the fluorescence intensity was measured using a Muse™ Cell Analyze. The percentages of apoptosis were examined using Multicaspase Kit (Millipore). After gathering cells, Multicaspase was added to each dish for 30 min at 37°C in dark and incubated with 7-aminoactinomycin D (7-AAD, Millipore) for 5 min at room temperature. The events for live, dead early, and late apoptotic cells were counted with the Muse Cell Analyzer.
6
Apoptosis and ROS in C2C12 Cells
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Using the methods described previously 11 (link), 28 (link), 29 (link), the apoptosis of C2C12 cells with or without THZ1 was measured by the Multicaspase Kit (Millipore). ROS was detected by the dihydroethidium (DHE; Millipore, EMD Millipore Corporation, Hayward,CA). We determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) method to measure cell viability.
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