Cell cycle and apoptosis were analyzed using propidium iodide (1 mg/ml) and ribonuclease-A (10 g/ml) (PI/RNase; BD Biosciences), and Annexin V/PI assay by flow cytometry (BD Biosciences, Franklin Lakes, NJ, USA) respectively as we previously described (37 (link)).
Annexin 5 pi assay
Manufactured by BD
Sourced in United States
The Annexin V/PI assay is a flow cytometry-based technique used to detect and quantify apoptosis in cell populations. Annexin V is a protein that binds to phosphatidylserine, which is exposed on the surface of apoptotic cells. Propidium iodide (PI) is a DNA-binding dye that can enter cells with compromised membranes, such as late apoptotic or necrotic cells. This assay allows for the identification of early apoptotic, late apoptotic, and necrotic cells in a sample.
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Lab products found in correlation
5 protocols using annexin 5 pi assay
1
Cell Cycle and Apoptosis Analysis
2
Annexin V/PI Apoptosis Assay
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Before exposure to the treatment conditions, cells were grown in 12-well plates until 70% confluence. Detection of apoptosis was done by Annexin V/PI Assay (BD Pharmingen, Bedford, MA, USA), according to the manufacturer's protocol. Briefly, cells were collected by trypsinization, washed twice with PBS and stained with 5 μl of Annexin V-fluorescein isothiocyanate and 5 μl of PI (5 μg/ml) in binding buffer for 15 min at room temperature in dark. The apoptotic cells were determined by FACSVerse (Model RUO 3L8C) flow cytometer (BD Biosciences).
3
Apoptosis Measurement by Flow Cytometry
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Briefly, after the indicated treatment, both floating and non-floating cells were collected and washed twice in PBS. The level of apoptosis was measured by flow cytometry using the Annexin V/PI assay (BD, Biosciences) according to the manufacturer's protocol using BD LSRII (BD, Biosciences) equipment. Data were collected on 10,000 cells. Apoptotic fractions were determined where cells positive for Annexin V staining but not propidium iodide were counted.
4
Inhibition of ILK in Cancer Cells
5
Cell Viability and Apoptosis Assay
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Cell viability was assessed with the CellTiter 96® AQueous One Solution Cell Proliferation (Promega, Cat# G3582) following manufacturer’s recommendations. Briefly, cells were seeded into 96-well plates at 3,000 cells/well in complete medium and incubated for 24 hr. Each drug was then added at various concentrations to triplicate wells in a final volume of 100 μL of medium. After 72 hr of incubation, 20 μL of MTS tetrazolium reagent was added, and the incubation was continued for an additional 2 hr at 37°C. The absorbance of soluble MTS tetrazolium formazan produced by viable cells following drug exposure was measured at 490 nm using a microplate reader. The sensitivity of cells to each drug was expressed as a percentage of the vehicle-treated control. For apoptosis assays, after the indicated treatment, both floating and non-floating cells were collected and washed twice in PBS. The level of apoptosis was measured by flow cytometry using the Annexin V/PI assay (BD, Biosciences, Cat#556547) according to the manufacturer’s protocol using BD LSRII (BD, Biosciences) equipment. Data were collected on 10,000 cells and analyzed with FlowJo software (FlowJo, RRID:SCR_008520).
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