The cell cycle was analyzed using the Cell Cycle Kit (BD, CA, USA). Cells were trypsinized and washed in phosphate buffered saline (PBS) for 5 min prior to collection by centrifugation at 1500 revolutions per minute (rpm). Cells were then fixed with 2 mL of prechilled 70% ethanol for 30 min at 4°C. The cells were then gently centrifuged and resuspended in a solution containing RNase A and PI. After 30 min incubations, the cells were analyzed by flow cytometry (BD Accuri C6, USA). For the apoptosis assay, the cells were trypsinized and washed with serum-containing medium. Cells were then centrifuged for 3 min at 1500 rpm and the supernatant was discarded. The cells were resuspended in 1 × Binding Buffer at 1–5 × 106/mL. The cells were then stained for 10–15 min at room temperature using the Annexin V-APC/PI Apoptosis Kit (eBioscience, CA, USA) in accordance with the manufacturer's instructions. The number of apoptotic cells was analyzed using flow cytometry (BD Accuri C6, USA).
Annexin 5 apc pi apoptosis kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Annexin V-APC/PI Apoptosis Kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide (PI), a DNA-binding dye, to differentiate between viable, early apoptotic, and late apoptotic/necrotic cells using flow cytometry or fluorescence microscopy.
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6 protocols using annexin 5 apc pi apoptosis kit
1
Cell Cycle and Apoptosis Analysis
2
Annexin V-APC/PI Apoptosis Assay
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Floating and adherent cells were collected and washed with PBS. Cells were then centrifuged for 2 minutes at 1500 rpm and resuspended in 1× Annexin‐V binding buffer at a concentration of 1 × 105 cells/mL. The cells were stained using the Annexin V‐APC/PI Apoptosis Kit (eBioscience, CA, USA) for 15 minutes at room temperature in the dark. Cells were analyzed by flow cytometry (BD Accuri C6, USA).
3
Annexin V-APC/PI Apoptosis Assay
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Floating and adherent cells were congregated and washed with phosphate-buffered saline (PBS). Cells were then centrifuged for 2 minutes at 1,500 rpm and resuspended in 1× Annexin-V binding buffer at a concentration of 1×105 cells/mL. The collected cells were stained with the Annexin V-APC/PI Apoptosis Kit (eBioscience, CA, USA) for 15 minutes at room temperature. The stained cells were analyzed by flow cytometry in accordance with the manufacturer’s protocols.
4
Cell Cycle and Apoptosis Analysis by Flow Cytometry
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Cells were collected at the logarithmic stage of growth, and were centrifuged and then resuspended at 2 × 106 cells/mL. After 30 min incubation in 80% ethanol at 4°C, propidium iodide (PI) staining was utilized to measure cell cycle stage using flow cytometry (BD Biosciences). The Annexin V-APC/PI Apoptosis Kit (eBioscience, San Diego, CA) was used for the apoptosis assay according to the manufacturer's instructions. Flow cytometery was performed to determine cell number, and data were analyzed using FlowJo9.1 software (Tree Star Inc., Ashland, OR).
5
Annexin V-APC/PI Apoptosis Assay
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The apoptosis rate was determined using an Annexin V-APC/PI apoptosis kit (eBioscience; Thermo Fisher Scientific, Inc.). GL261 cells were treated with HSP at indicated concentrations for 24 h. The cells were then harvested and stained with Annexin V-APC and propidium iodide (PI). The apoptosis rate was calculated using a flow cytometer and FlowJo software (version 10; BD, Franklin Lakes, NJ, USA).
6
Annexin V-APC/PI Apoptosis Assay
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The apoptotic status was analyzed by using an Annexin V-APC/PI Apoptosis Kit (eBioscience, CA, USA) in accordance with the manufacturer’s instructions. Briefly, cells (1 × 106cells/mL) were collected by EDTA treatment and incubated with a mixture of annexin V-FITC and PI for 15 min at room temperature. The number of apoptotic cells was analyzed by flow cytometry (BD Accuri C6, USA).
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