AIF (apoptosis-inducing factor) antibody, NAS, and 2,7-dichlorodihydrofluorescein diacetate were obtained from Sigma (St. Louis, MO). Cleaved caspase-3, cleaved caspase-9, Cyt-C, and β-actin antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA). Cox-IV antibodies came from BD PharMingen (San Diego, CA). Horseradish-conjugated secondary anti-mouse antibody and anti-rabbit antibody were purchased from Amersham Pharmacia Biotech (Piscataway, NJ). FITC-conjugated secondary antibodies were purchased from Jackson Immunoresearch (West Grove, PA). Hoechst 33342, rhodamine 123, and tetramethylrhodamine methyl ester (TMRM) were purchased from Life Technologies Corporation (Grand Island, NY). The enhanced chemiluminescence (ECL) system was purchased from Amersham Pharmacia Biotech (Piscataway, NJ). MDA and SOD kits were obtained from the NanJing JianCheng Bioengineering Institute (Nanjing, China). The TUNEL apoptosis kits came from Boster Biotechnology (Wuhan, China). Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit was purchased from Roche (Germany).
Annexin 5 fitc propidium iodide apoptosis detection kit
Manufactured by Roche
Sourced in Germany
The Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit is a laboratory tool used to detect and quantify apoptosis, a programmed cell death process. The kit utilizes Annexin V, a calcium-dependent phospholipid-binding protein, conjugated with the fluorescent dye FITC, and propidium iodide, a DNA-binding dye. This combination allows for the identification of cells undergoing early and late stages of apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Sod and mda kit, by Nanjing Jiancheng (1 mentions)
Fitc conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Rhodamine 123, by Thermo Fisher Scientific (1 mentions)
Tetramethylrhodamine methyl ester tmrm, by Thermo Fisher Scientific (1 mentions)
2 7 dichlorodihydrofluorescein diacetate, by Merck Group (1 mentions)
Cleaved caspase 9, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Cyt c, by Cell Signaling Technology (1 mentions)
Facscalibur, by BD (1 mentions)
Hoechst 33258, by Beyotime (1 mentions)
2 protocols using annexin 5 fitc propidium iodide apoptosis detection kit
1
Apoptosis Signaling Pathway Characterization
2
Apoptosis and Cell Cycle Analysis in CRC Cells
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Morphological characteristics of apoptotic cells were observed by Hoechst 33258 staining. CRC cells infected with VG9-IL-24, VG9-EGFP, or PBS for 24 h were incubated with Hoechst 33258 (Beyotime Biotechnology, Shanghai, China) for 30 min. The apoptotic morphological changes of cells were observed under an Olympus IX51 fluorescence microscope (Olympus Corporation, Tokyo, Japan) immediately.
Apoptosis was further quantified by flow cytometric analysis using the Annexin-V–FITC/Propidium Iodide (PI) Apoptosis Detection kit (Roche Applied Science, Penzburg, Germany). HCT116 cells were infected with VG9-IL-24, VG9-EGFP, or PBS for 48 h, and apoptotic cells were detected according to the manufacturer’s instructions using BD FACSCalibur (BD Biosciences, Mountain View, CA, USA).
For cell cycle detection, HCT116 cells seeded in six-well plates were infected with VG9-IL-24, VG9-EGFP, or PBS for 48 h and then fixed in 70% cold ethanol overnight at −20°C. Cells were washed with PBS and resuspended in 50 μg/ml of PI solution. After incubation for 30 min in the dark at 37°C, the treated cells were analyzed by flow cytometry (BD FACSCalibur, BD Biosciences). The percentages of G0/G1, S, and G2/M stage cells were quantified using Flow Jo Software (Tristar, Mountain View, CA, USA).
Apoptosis was further quantified by flow cytometric analysis using the Annexin-V–FITC/Propidium Iodide (PI) Apoptosis Detection kit (Roche Applied Science, Penzburg, Germany). HCT116 cells were infected with VG9-IL-24, VG9-EGFP, or PBS for 48 h, and apoptotic cells were detected according to the manufacturer’s instructions using BD FACSCalibur (BD Biosciences, Mountain View, CA, USA).
For cell cycle detection, HCT116 cells seeded in six-well plates were infected with VG9-IL-24, VG9-EGFP, or PBS for 48 h and then fixed in 70% cold ethanol overnight at −20°C. Cells were washed with PBS and resuspended in 50 μg/ml of PI solution. After incubation for 30 min in the dark at 37°C, the treated cells were analyzed by flow cytometry (BD FACSCalibur, BD Biosciences). The percentages of G0/G1, S, and G2/M stage cells were quantified using Flow Jo Software (Tristar, Mountain View, CA, USA).
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