To assess cell viability, cells were washed once with PBS and stained with 1 µg/mL propidium iodide (PI, Molecular Probes) for 10 minutes at room temperature. To assess apoptotic cell death, cells were washed once with PBS and stained with PI and Annexin-V (eBioscience) and processed according to the manufacturer’s protocol. Flow analysis was carried out using the 9-color CyAn™ ADP from Beckman Coulter (Miami, FL). To assess cell viability by Trypan Blue exclusion, cells were washed once with PBS and stained with Trypan Blue solution (Sigma). Cell viability was determined using TC20 automated cell counter (BioRad).
Annexin 5 and pi
Manufactured by Thermo Fisher Scientific
Sourced in United States
Annexin V and PI are laboratory reagents used for cell analysis. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine (PS), which is exposed on the surface of apoptotic cells. Propidium iodide (PI) is a fluorescent dye that binds to DNA and is commonly used to detect necrotic or late-stage apoptotic cells. These reagents are often used together in flow cytometry and other cell analysis techniques to identify and quantify different cell populations based on their stage of cell death.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Thermo Fisher Scientific (2 mentions)
Cyan adp 9 color, by Beckman Coulter (2 mentions)
Trypan blue solution, by Merck Group (2 mentions)
Tc20 automated cell counter, by Bio-Rad (2 mentions)
Calibur, by BD (2 mentions)
Facscelesta, by BD (1 mentions)
Facscanto 2, by BD (1 mentions)
Cd86 pe, by Thermo Fisher Scientific (1 mentions)
Cd4 fluorescein isothiocyanate, by Thermo Fisher Scientific (1 mentions)
Cd8 pe, by Thermo Fisher Scientific (1 mentions)
Cd3 percp cyanine 5, by Thermo Fisher Scientific (1 mentions)
Cd11c apc, by Thermo Fisher Scientific (1 mentions)
Cd4 fitc, by Thermo Fisher Scientific (1 mentions)
Dead cell apoptosis detection kit, by Thermo Fisher Scientific (1 mentions)
Efluor 670, by Thermo Fisher Scientific (1 mentions)
Tunel apoptosis detection kit, by Boster Bio (1 mentions)
13 protocols using annexin 5 and pi
1
Cell Viability and Apoptosis Assessment
2
Cell Viability and Apoptosis Assessment
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To assess cell viability, cells were washed once with PBS and stained with 1 µg/mL propidium iodide (PI, Molecular Probes) for 10 minutes at room temperature. To assess apoptotic cell death, cells were washed once with PBS and stained with PI and Annexin-V (eBioscience) and processed according to the manufacturer’s protocol. Flow analysis was carried out using the 9-color CyAn™ ADP from Beckman Coulter (Miami, FL). To assess cell viability by Trypan Blue exclusion, cells were washed once with PBS and stained with Trypan Blue solution (Sigma). Cell viability was determined using TC20 automated cell counter (BioRad).
3
Annexin V and PI Staining
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After the described treatment, cells were collected and washed twice with prewarmed FBS-free DMEM and then stained with PI and Annexin V (Thermo Fisher) for 20 min at 37°C. After staining, cells were washed twice with prewarmed PBS for analysis with a flow cytometer (BD Calibur).
4
Apoptosis Analysis by Flow Cytometry
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After the described treatment, cells were collected and washed twice with prewarmed FBSfree DMEM and then stained with PI and Annexin V (Thermo Fisher) for 20 min at 37 °C. After staining, cells were washed twice with prewarmed PBS for analysis with a flow cytometer (BD Calibur).
5
Quantifying Myoblast Apoptosis by Flow
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Primary myoblasts were collected using trypsin, counted, and washed in ice-cold PBS before resuspension in Annexin V buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4). Cells were labeled with Annexin V and PI (Life Technologies) and analyzed by flow cytometry on a BD FACSCelesta instrument (BD Biosciences), and dot plots were made and analyzed using Kaluza v1.2 software (Brea).
6
Photodynamic Therapy Cytotoxicity Assay
7
Apoptosis Assay with Annexin V and PI
8
Cisplatin-Induced Apoptosis Assay
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siRNA-treated cells or stable transfectants of shCISD2 were seeded into six-well plates and incubated overnight. Culture media were replaced by a serum-free medium with or without cisplatin (30 μM), and cells were harvested at 12, 24 and 48 h, labeled with annexin V and PI (Invitrogen) per the manufacturer’s instructions, and then analyzed by flow cytometry.
9
Splenic Lymphocyte Immunophenotyping Protocol
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Splenic lymphocytes were centrifuged at 180×g for 10 min and the cells were collected. The cells were re-suspended in PBS to a density of 2×105 cells/ml, and labelled with anti-mouse monoclonal antibodies, including CD3-PerCP-Cyanine5.5 (45-0031-82), CD4-fluorescein isothiocyanate (CD4-FITC; 11-0041-82), CD8-phycoerythrin (CD8-PE; 12-0081-81), CD11c-APC (17-0114-82), major histocompatibility complex (MHC) class II-FITC (11-5322-81), CD86-PE (12-0869-42), CD40-PE-Cyanine7 (25-0409-42), and IDO-APC (17-9477-41) (eBioscience, San Diego, CA, USA), and Annexin-V and PI (V13241; Invitrogen, Shanghai, China). For CD4 and CD8 T cell staining, cells were incubated with 1 µl of CD3-PerCP-Cyanine5.5, 2 µl of CD4-FITC, and 2 µl of CD8-PE for 25 min at room temperature in the dark. Apoptosis was evaluated by incubation with Annexin-V and PI for 30 min at room temperature. For DCs markers, cells were incubated with 1 µl of CD11c-APC, 2 µl of MHC class II-FITC, 2 µl of CD86-PE, and 2 µl of CD40-PE-Cyanine7 for 30 min at room temperature. Samples were detected using an LSR II (BD Biosciences, NJ, USA) flow cytometer and analyzed using the FlowJo software (TreeStar, NJ, USA) according to the manufacturer’s instructions.
10
Apoptosis Detection by Flow Cytometry
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Flow cytometry was used to detect the percentage of apoptotic cells by a Dead Cell Apoptosis Detection Kit with Annexin V and PI (Invitrogen). Briefly, at 48 h after transfection of plasmid, cells were harvested and suspended in Annexin Binding buffer. After that, PI and Annexin V were added into the above cell suspension and incubated at room temperature for 30 min. Afterwards, cells were subjected to flow cytometry for analysis of apoptosis. The cells positive for Annexin V was at early apoptosis and cells positive for both Annexin V and PI were at later apoptosis.
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