Human primary cardiac‐derived microvascular endothelial cells were incubated with varying MV.Act concentrations for different time periods, processed with MitoSOX™ Red Mitochondrial Superoxide Indicator (Thermofisher, M36008) and analysed by flow cytometry. Apoptosis was assessed using Annexin V detection kit (BioLegend).
Annexin 5 detection kit
Manufactured by BioLegend
Sourced in United States
The Annexin V detection kit is a laboratory tool used to detect and quantify the presence of Annexin V, a protein that binds to phosphatidylserine. Annexin V is commonly used as a marker for apoptosis (programmed cell death) in various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Mitosox red mitochondrial superoxide indicator, by Thermo Fisher Scientific (1 mentions)
Anti cd105, by BioLegend (1 mentions)
Anti cd14 igg, by BioLegend (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Brdu elisa kit, by Roche (1 mentions)
Aria 3 system, by BD (1 mentions)
Facscelesta, by BD (1 mentions)
5 protocols using annexin 5 detection kit
1
Cardiac Microvascular Endothelial Cell Responses
2
Comprehensive Characterization of MSCs
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Flow cytometry was performed to assess protein surface marker expression. Isolated MSCs were washed and incubated with respective MSC surface marker antibodies (2 ug/ml anti‐CD45 IgG, anti‐CD34 IgG, anti‐CD14 IgG, anti‐CD44 IgG, anti‐CD90 IgG and anti‐CD105) or isotype control purchased from BioLegend (San Diego, CA), followed by flow cytometry analysis on a FACS Calibur flow cytometer (BD Biosciences). Senescence was determined with intracellular anti‐pSTAT3 staining (BioLegend). Apoptosis was determined with an Annexin V detection kit (BioLegend) and intracellular pSTAT3 staining according to manufacturer's protocol. Proliferation assay was performed with a BrdU ELISA kit (Roche) according to manufacturer's protocol.
3
Imaging Flow Cytometric Analysis of Phosphatidylserine
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Cell or MV suspensions labelled with fluorescent dyes or antibodies were analysed using Amnis® Mark II ImageStreamX imaging flow cytometer (Merck Millipore).12 Cells were FACsorted using an Aria III system (BD Bioscience). Phosphatidylserine staining was performed using Annexin V detection kit (BioLegend).
4
Apoptotic Effects of Compounds 2 and 4 on HL-60 Cells
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The apoptotic effects
of compound 2 and compound 4 on HL-60 cells were investigated using
the annexin V detection kit (BioLegend) in accordance with the manufacturer’s
instructions. The human promyelocytic leukemia cell line was plated
in 6-well plates at a density of 1.2 × 105 cells/well.
The cells were exposed to the active substances (2 and 4 ) at an IC50 concentration for 24 h at 37 °C
and 5% CO2. Next, the cells were centrifuged at 200 rmp
and washed twice with PBS buffer. The pellets were resuspended and
stained with propidium iodide and annexin V in annexin binding buffer
(ABB). After double staining, the cells were incubated in the dark
for 30 min and analyzed in a flow cytometry analyzer (BD FACSCelesta).15 (link)
of compound 2 and compound 4 on HL-60 cells were investigated using
the annexin V detection kit (BioLegend) in accordance with the manufacturer’s
instructions. The human promyelocytic leukemia cell line was plated
in 6-well plates at a density of 1.2 × 105 cells/well.
The cells were exposed to the active substances (
and 5% CO2. Next, the cells were centrifuged at 200 rmp
and washed twice with PBS buffer. The pellets were resuspended and
stained with propidium iodide and annexin V in annexin binding buffer
(ABB). After double staining, the cells were incubated in the dark
for 30 min and analyzed in a flow cytometry analyzer (BD FACSCelesta).15 (link)
5
Apoptotic Effects of Garlic SEVs on Cancer Cells
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The apoptotic effect of garlic SEVs on cancer cells was detected with Annexin V Detection Kit (BioLegend, USA) according to the manufacturers’ protocol. A498, A549 and HDF cells were seeded onto 6-well plates with the cell density of 1 × 105 and treated with 50 μg/ml of A. Sativum SEVs for 24 and 48 h. Following the incubation process, cells harvested were incubated with Annexin V FITC and PI PE conjugated antibody and Apoptosis and necrosis were evaluated using flow cytometer. Obtained data was then analyzed using CellQuest Pro program.
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