The percentage of early-stage apoptosis and lately-stage apoptosis, necrosis of cells induced by the addition of LPS or LPS/PEs mixture were determined through Annexin V-FITC/PI double staining assay kit (Beyotime). Cells were seeded and treated with PEs solutions together with 1 µg/mL of LPS or LPS alone. Nontreatment wells were as controls. After incubation of PEs mixed with LPS for 24 h, suspended medium (200 µL) and trypsin-disassociated cells were collected and centrifuged. The precipitate was resuspended in 195 µL of binding buffer plus 5 µL of Annexin V-FITC for 5-min incubation followed by 10-min incubation of 10 µL Propidium Iodide (PI). The nonstaining sample was as a reference. After a gentle mix at room temperature and a dark environment was conducted, the percentage of cell death was detected via Flow Cytometry (FCM). The cell plot was distributed into four quadrants based on Annexin V-FITC and PI axes representing living cells in quadrant 3, early-stage apoptotic cells in quadrant 4, and total lately-stage apoptotic and necrotic cells in quadrant 1, respectively.
Annexin 5 fitc pi double staining assay kit
Manufactured by Beyotime
Sourced in China
The Annexin V-FITC/PI double staining assay kit is a laboratory tool used to detect and quantify apoptosis, a form of programmed cell death. The kit contains Annexin V-FITC, a fluorescent dye that binds to phosphatidylserine, and propidium iodide (PI), a dye that stains DNA. This combination allows for the differentiation of viable, early apoptotic, and late apoptotic/necrotic cells through flow cytometry analysis.
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Lab products found in correlation
2 protocols using annexin 5 fitc pi double staining assay kit
1
Quantifying Cell Death Mechanisms
2
Anticancer Potential of Shikonin Compounds
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The chemical reagents used in our study including Acetylshikonin (lot number: A832580, 99% purity), Shikonin (lot number: S914687, 98% purity), Deoxyshikonin (lot number: D861265, 98% purity), β,β-dimethylacrylshikonin (lot number: D799242, 98% purity), Lithospermidin E, Shikonofuran A (lot number: C11637623, 99% purity) were all purchased from MACKLIN (Shanghai, China). Cell counting kit-8 (CCK-8) assay kit, DNA content assay kit (Solibao, Beijing, China) were obtained from Solibao (Beijing, China). Annexin V-FITC/PI double staining assay kit, caspase-3 enzyme activity kit, JC-1 kit wer purchased from Beyotime (Shanghai, China). Other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise specified.
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