The Annexin V‐fluorescein isothiocyanate propidium iodide (FITC/PI) apoptosis detection kit (KeyGEN BioTECH Co. Ltd., Nanjing, China) was purchased to detect cell apoptotic rate, in light of manufacturer’s instruction. In brief, cells were collected into flow tubes and rinsed with PBS. After cells were suspended with binding buffer (500 µL), they were cultivated with Annexin V/FTIC (5 µL) and PI (5 µL) at room temperature for 15 min darkly. Cell apoptotic rate was calculated using a FACScan flow cytometer (BD Biosciences).
Annexin 5 fluorescein isothiocyanate propidium iodide fitc pi apoptosis detection kit
Manufactured by Keygen Biotech
Sourced in China, United States
The Annexin V-fluorescein isothiocyanate propidium iodide (FITC/PI) apoptosis detection kit is a laboratory tool used to detect and quantify apoptosis, a programmed cell death process. The kit utilizes the binding properties of Annexin V, a protein that binds to phosphatidylserine, and the DNA-binding dye propidium iodide to identify cells undergoing early and late stages of apoptosis.
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3 protocols using annexin 5 fluorescein isothiocyanate propidium iodide fitc pi apoptosis detection kit
1
Annexin V-FITC/PI Apoptosis Detection
2
Cytotoxicity and Apoptosis Assay Protocol
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Reagents purchased for the present study were 0.25% trypsin-0.02% EDTA (Jinuo Biomedical Technology Co., Ltd., Hangzhou, China), fetal bovine serum (FBS), Roswell Park Memorial Institute (RPMI) 1640 medium (both from Hangzhou Sijiqing Biological Engineering Co., Ltd., Hangzhou, China), an MTT assay kit (Amresco LLC, Solon, OH, USA), an Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI)apoptosis detection kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China). All other reagents were manufactured domestically and analytically pure.
3
Apoptosis and Necrosis Analysis in SGC-7901 Cells
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SGC-7901 cells were treated with MK-2206 at concentrations ranging from 0 to 16 μM for 72 hours. An Annexin-V–fluorescein isothiocyanate/propidium iodide (FITC/PI) apoptosis detection kit (KeyGEN Biotech, Nanjing, People’s Republic of China) was used to measure apoptosis and necrosis in drug-treated cells according to the manufacturer’s instruction, using an FACScan flow cytometer (BD Biosciences, San Jose, CA, USA). Viable cells were negative for both Annexin-V and PI (Annexin-V−/PI−), while late-stage apoptotic cells and necrotic cells were double-stained with Annexin-V and PI (Annexin-V+/PI+) and apoptotic cells were stained positive only with Annexin-V (Annexin-V+/PI−).
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