Apoptosis analysis was performed by staining with Annexin V/7-AAD, using the Muse™ Cell Analyzer (Merck Millipore, Milan, Italy) in according to the manufacturer's instructions. In brief, a 100 μl treated cell suspension was labeled for 20 min in the dark with the same volume of the MuseTM Annexin-V & Dead Cell reagent (Merck Millipore). Subsequently, quantitative detection of Annexin-V/7-AAD positive cells was performed using the Muse™ Cell Analyzer.
Muse annexin 5 dead cell reagent
Manufactured by Merck Group
Sourced in United States, Germany
The Muse™ Annexin V & Dead Cell Assay Reagent is a fluorescent reagent used for the detection and quantification of apoptotic and dead cells in samples. It contains Annexin V conjugated to a fluorescent dye and a dead cell stain.
Automatically generated - may contain errors
Lab products found in correlation
Muse cell analyzer, by Merck Group (24 mentions)
Muse annexin 5 dead cell kit, by Merck Group (3 mentions)
Dulbecco s modified eagle s medium dmem, by PAN Biotech (3 mentions)
Fetal bovine serum (fbs), by PAN Biotech (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Muse 1.4 analysis software, by Merck Group (1 mentions)
Muse cell analyzer flow cytometry, by Merck Group (1 mentions)
Muse cell analyzed flow cytometry, by Merck Group (1 mentions)
Muse count viability reagent, by Merck Group (1 mentions)
Muse cell analyser, by Merck Group (1 mentions)
Muse caspase 3 7 kit, by Merck Group (1 mentions)
Muse 1.5 analysis software, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Accutase, by Merck Group (1 mentions)
Facsaria, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Gossypol, by Merck Group (1 mentions)
Whole cell lysis buffer, by iNtRON Biotechnology (1 mentions)
Antibiotic antimycotic 100, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
34 protocols using muse annexin 5 dead cell reagent
1
Apoptosis Analysis by Annexin V/7-AAD
2
Annexin V/7-AAD Apoptosis Assay
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Apoptosis analysis was performed by staining with Annexin V/7-AAD, using the MuseTM Cell Analyzer in according to the manufacturer’s instructions. In brief, a 100 μl treated cell suspension was labeled for 20 min in the dark with the same volume of the MuseTM Annexin-V & Dead Cell reagent (Merck Millipore). Subsequently, quantitative detection of Annexin-V/7-AAD positive cells was performed with the MuseTM Cell Analyzer [51 (link)].
3
Apoptosis Induction in Gastric Cancer
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Annexin V and Propidium Iodide (PI) staining assay was carried out using the method given by Kloesch et al., with minor modifications [15] . Briefly, to confirm the apoptosis induced by Niclosamide on HGC-27 and MKN-74, cells in a six-well plate were treated with 0.5 µM,1 µM, 2 µM and 2.5 µM, 5 µM, 10 µM concentrations of Niclosamide in HGC-27 and MKN-74 cell lines respectively for 48hrs. Then cells were treated with MuseTM Annexin-V & Dead Cell reagent (Merck-Millipore, Germany) as per instructions provided by the manufacturer and analyzed by Muse™ Cell Analyzer (Merck-Millipore, Germany).
4
Apoptosis Assay of Melanoma Cells
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1205Lu, UACC903, and SK-Mel-8 (4 × 105 cells/well) were pleated and treated with DMSO (control) or compound 1 at the given concentrations. After 24 h of treatment, both floating and attached cells were collected and stained with the MuseTM Annexin V & Dead Cell Reagent (Millipore; Bedford, MA, USA) for 20 min at room temperature in the dark. The results were collected by MuseTM Cell Analyzer (Millipore).
5
Apoptosis and Necrosis Assessment
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Cells were incubated with MuseTM Annexin V & Dead Cell Reagent (Millipore) for 20 min at RT in the dark. Apoptotic and necrotic cell analysis was performed using a MuseTM Annexin V & Dead Cell Kit (Millipore) and MuseTM Cell Analyzer (Millipore).
6
Quantifying Apoptosis after Drug Treatment
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In order to determine the number of apoptotic cells after drug treatment, Muse Annexin V & Dead Cell Assay (Sigma-Aldrich) was performed following the manufacturer’s protocol. 200,000 cells/well were seeded in 6-well plates. After cell cultures reached 70%–80% of confluence, they were treated with the drugs as described in the Results section. Adherent cells and cells in the suspension were harvested and diluted in PBS. 100 μl of cell suspension was mixed with 100 μl Muse™ Annexin V & Dead Cell Reagent, incubated for 20 min, and their apoptotic profile was analysed through Muse™ Cell Analyzer (Merck Millipore, Darmstadt, Germany).
7
Annexin V Apoptosis Assay
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Cells were seeded on 6-well overnight and then treated with different concentrations of AA for 24 h. Cells were harvested and suspended in PBS (2% (v/v) BSA), and then incubated with Muse™ Annexin V & Dead Cell reagent (EMD Millipore, Billerica, MA, USA) for 20 min at room temperature in dark. Samples were analyzed by Muse Cell Analyzer flow cytometry (EMD Millipore, Billerica, MA, USA) and by MUSE 1.4 Analysis software (EMD Millipore).
8
Measuring Apoptosis Rate via Annexin V
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The apoptosis rate of the cells was confirmed by using a Muse™ Annexin V & Dead Cell kit (Merck Millipore) according to the manufacturer’s protocols. In brief, the cells were washed with PBS and resuspended in 100 µL of Muse™ Annexin V & Dead Cell reagent (Merck Millipore) for 20 min at room temperature (RT) in the dark. The dead cells were analyzed using a Muse™ Cell Analyzer (Merck Millipore).
9
Apoptosis Induction by 7-Epitaxol
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As previously described,21 the cell lines were treated with different concentrations of 7‐Epitaxol for 24 h. Then, the cells were harvested and suspended in PBS (2% BSA) and incubated with Muse Annexin V & Dead Cell reagent (EMD Millipore, Billerica, MA) for 20 min at room temperature in dark. The data were analysed by Muse Cell Analyzed flow cytometry (Merck Millipore, Burlington, MA).
10
Apoptosis Assay for A549 Cells
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For the apoptosis assay, A549 cells
were seeded at a density of 5 × 104 cells/well in
24-well plates and incubated overnight at 37 °C in 5% CO2. Then, the cells were treated for 24 h with solvent 10.8,
108.1, or 1081 nM brusatol or solvent control diluted in culture medium.
After removing the compound-containing medium, the cells were trypsinized.
Next, the cell samples were incubated for 20 min at room temperature
with the Muse Annexin V & Dead Cell reagent (Merck Millipore Burlington,
USA) that uses Annexin V as a marker for apoptosis and 7-aminoactinomycin
D as a marker for dead and necrotic cells. After staining with these
markers, cells were analyzed according to the manufacturer’s
instructions by flow cytometry. For analysis, the percentage of viable,
early apoptotic, late apoptotic, and necrotic cells in relation to
the total number of cells was determined, and the sum of early and
late apoptotic cells was used to calculate the percentage of total
apoptotic cells.
were seeded at a density of 5 × 104 cells/well in
24-well plates and incubated overnight at 37 °C in 5% CO2. Then, the cells were treated for 24 h with solvent 10.8,
108.1, or 1081 nM brusatol or solvent control diluted in culture medium.
After removing the compound-containing medium, the cells were trypsinized.
Next, the cell samples were incubated for 20 min at room temperature
with the Muse Annexin V & Dead Cell reagent (Merck Millipore Burlington,
USA) that uses Annexin V as a marker for apoptosis and 7-aminoactinomycin
D as a marker for dead and necrotic cells. After staining with these
markers, cells were analyzed according to the manufacturer’s
instructions by flow cytometry. For analysis, the percentage of viable,
early apoptotic, late apoptotic, and necrotic cells in relation to
the total number of cells was determined, and the sum of early and
late apoptotic cells was used to calculate the percentage of total
apoptotic cells.
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