5×102 PyMT or 4T1 cells and 2×103 EO771 cells were plated for 24 hours before treatment with rDKK1 (50, 100, 200 ng/ml) or tamoxifen (1, 2, 10 μM) for indicated amount of time. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Invitrogen) and annexin V assay (ThermoFisher Scientific) were performed per manufacturer’s protocol.
Annexin 5 assay
Manufactured by Thermo Fisher Scientific
Sourced in United States
Annexin V assay is a laboratory technique used to detect and quantify apoptosis, a programmed cell death process. The assay utilizes Annexin V, a calcium-dependent phospholipid-binding protein, which binds to phosphatidylserine exposed on the surface of apoptotic cells. This binding can be measured using various detection methods, such as flow cytometry or fluorescence microscopy, to determine the extent of apoptosis in a sample.
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Lab products found in correlation
4 protocols using annexin 5 assay
1
Evaluating Cell Proliferation and Apoptosis
2
Cell Sorting and Cycle Analysis
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Cells were dissociated using trypsin and transferred into PBS supplemented with 3% FCS. FACSAria IIu sorter (BD biosciences) was used for sorting and analysis. Cell cycle analysis using EdU assay (Thermo) and cell death analysis using Annexin V assay (Thermo) were performed according the manufacturer’s instructions. Flowjo software (TreeStar) was used for data analysis.
3
Apoptosis Detection by Annexin V Assay
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An annexin V assay was performed according to the manufacturer’s instructions (Life Technologies, Carlsbad, CA, USA). Nobiletin- and/or palbociclib-treated cells were collected, and used for annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. The samples were then analyzed using flow cytometry within 1 h. Each experiment was conducted in triplicate.
4
Annexin V Apoptosis Assay Protocol
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An Annexin V assay was performed according to the manufacturer's instructions (Life Technologies, Carlsbad, CA, USA). Nobiletin and/or palbociclib treated cells were collected, and used for Annexin V-FITC/PI staining. After that, the samples analyzed via ow cytometry within 1 h. Each experiment was conducted in triplicate.
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