AGS and MKN45 were seeded into 6-well plates at a density of 1 × 106 cells per well and cultured for 24 h. Then the cells were washed with PBS three times, trypsinised, and harvested for staining using an annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (Keygen, Nanjing, China). The staining of cells was assessed by flow cytometry and data analysed using the FlowJo software, with FITC-positive cells considered apoptotic.
Annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis kit
Manufactured by Keygen Biotech
Sourced in China
The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit is a laboratory tool used to detect and quantify apoptosis, a programmed cell death process. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and PI, a DNA-binding dye, to distinguish between viable, early apoptotic, and late apoptotic/necrotic cells.
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6 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis kit
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Annexin V-FITC/PI Apoptosis Assay
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Annexin V-FITC/PI Apoptosis Assay
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The proportion of apoptotic cells were determined by using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China) following the manufacturers instructions. Cells cultured in 6-well plates were transfected with siTRIM11 or siNC. After incubating for 48 h, cells were harvested, labeled with Annexin V-FITC and PI, and analyzed using a FACScan flow cytometry (BD Biosciences, San Jose, CA, USA) within 1 h.
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Apoptosis and Cell Proliferation Assays
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Cells were seeded into 6-well plates at a density of 1×106 cells per well and cultured for 24 hours. Cells were then treated with either fresh DMEM or 8 μg/mL DOX. After incubation for 24 hours, cells were stained with an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (Keygen, Nanjing, China). The staining of cells was assessed by flow cytometry.
In addition, to evaluate the interaction of miR-222-3p and FOXP2, the LoVo/ADR cells infected with miR-222-3p inhibitors were again treated with siRNA-FOXP2, before being analyzed by CCK8, EdU assay, and flow cytometry as described.
In addition, to evaluate the interaction of miR-222-3p and FOXP2, the LoVo/ADR cells infected with miR-222-3p inhibitors were again treated with siRNA-FOXP2, before being analyzed by CCK8, EdU assay, and flow cytometry as described.
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Apoptosis Assay in Liver Cancer Cells
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The Huh7 and HepG2 liver cancer cells were collected after digestion by trypsin enzyme. Subsequently, the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (KeyGen, China) was used to stain the cells. Then, the collected cells were analyzed with a FACS Canto II flow cytometer (BD Biosciences).
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Apoptosis Detection with Hoechst 33342 and Annexin V-FITC/PI
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Hoechst 33342 (Sigma-Aldrich, St. Louis, Missouri, USA), Annexin V- fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (Keygen, Nanjing, China).
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Annexin V-FITC/PI Apoptosis Assay
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Cell apoptosis was determined using the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (KeyGen Biotechnology, Nanjing, China) according to the manufacturer’s instructions. In brief, MCF-7 cells in different groups were seeded onto six-well plates with 40,000 cells per well and cultured overnight. The next day, cells were trypsinized, washed with ice-cold PBS, and fixed with 75% ethanol. The fixed cells were incubated with 5 μL Annexin V-FITC and 5 μL PI for 48 h. All samples were harvested, and the apoptotic rate was assessed by flow cytometry (BD Biosciences, USA). This experiment was performed three times independently.
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