Apoptosis analysis was conducted as previously described.24 (link) Exponentially-growing HUVECs were collected and stained with an Annexin V-FITC/PI Assay Kit (BestBio, Shanghai, China) in accordance with the manufacturer's protocol. The apoptosis detection included three groups: 3-h ischemia group (group I), 3-h ischemia/24-h reperfusion group (group IR), and 3-h normal control group (group NC). The cells were then subjected to flow cytometry analysis using a BD Influx (BD Bioscience, San Jose, CA, USA). The acquired flow cytometry data were analyzed with FlowJo v10.0 software (Becton, Dickinson and Company, Franklin Lakes, NJ, USA). The assay was performed in triplicate under each condition.
Annexin 5 fitc pi assay kit
Manufactured by BestBio
Sourced in China
The Annexin V-FITC/PI Assay Kit is a laboratory reagent used for the detection and quantification of apoptosis in cells. It utilizes Annexin V, a calcium-dependent phospholipid-binding protein, conjugated with the fluorescent dye FITC, and the nuclear stain propidium iodide (PI) to identify cells undergoing early and late apoptosis.
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Lab products found in correlation
3 protocols using annexin 5 fitc pi assay kit
1
Apoptosis Analysis in HUVEC Cells
2
Apoptosis and CD133 Expression Analysis
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Cell apoptosis was detected by flow cytometry analysis using Annexin V-FITC/PI Assay kit (BB-4101, BestBio, Shanghai, China) following the instruction of manufacturer. Briefly, after 48 h post-irradiation (0 or 4 Gy), the cells were digested with trypsin (without EDTA), washed with PBS, and then stained with annexin V-FITC (5 µl) and PI (5 µl). The apoptotic cells were analyzed by flow cytometry (BD Biosciences, San Diego, USA) and Beckman Flow Cytometry Analyzer (Beckman CytoFLEX, CA, USA).
The CD133 expression level was also detected by flow cytometry analysis. The cells were digested with trypsin (without EDTA), and stained with the human anti-CD133/FITC (bs-0395R-FITC, Bioss, Beijing, China). The CD133 expression was analyzed by flow cytometry (BD Biosciences, San Diego, USA) and Beckman Flow Cytometry Analyzer (Beckman CytoFLEX, CA, USA). Each experiment was performed in triplicate.
The CD133 expression level was also detected by flow cytometry analysis. The cells were digested with trypsin (without EDTA), and stained with the human anti-CD133/FITC (bs-0395R-FITC, Bioss, Beijing, China). The CD133 expression was analyzed by flow cytometry (BD Biosciences, San Diego, USA) and Beckman Flow Cytometry Analyzer (Beckman CytoFLEX, CA, USA). Each experiment was performed in triplicate.
3
Ischemia-Induced HUVEC Apoptosis Analysis
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The establishment of a lethal ischemic
model of HUVECs and apoptosis analysis was conducted as previously
described.22 (link) This analysis included three
groups: a 3 h ischemia group (referred to as group I), a 3 h ischemia/24
h reperfusion group (referred to as group IR), and a 3 h normal control
group (referred to as group NC). The exponentially growing HUVECs
of these three groups were collected and stained with an Annexin V-FITC/PI
assay kit (BestBio, Shanghai, China) following the manufacturer’s
protocol. Then, cells were subject to flow cytometry analysis using
BD influx (BD Biosciences, San Jose, CA, USA). The acquired flow cytometry
data were analyzed with FlowJo v10.0 software (Becton, Dickinson and
Company, Franklin Lakes, NJ, USA). Each assay condition was done in
triplicate.
model of HUVECs and apoptosis analysis was conducted as previously
described.22 (link) This analysis included three
groups: a 3 h ischemia group (referred to as group I), a 3 h ischemia/24
h reperfusion group (referred to as group IR), and a 3 h normal control
group (referred to as group NC). The exponentially growing HUVECs
of these three groups were collected and stained with an Annexin V-FITC/PI
assay kit (BestBio, Shanghai, China) following the manufacturer’s
protocol. Then, cells were subject to flow cytometry analysis using
BD influx (BD Biosciences, San Jose, CA, USA). The acquired flow cytometry
data were analyzed with FlowJo v10.0 software (Becton, Dickinson and
Company, Franklin Lakes, NJ, USA). Each assay condition was done in
triplicate.
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