Cellular apoptosis was measured using the Annexin V fluorescein isothiocyanate (FITC)‐propidium iodide (PI) staining kit (Sigma) according to the manufacturer's protocol. Briefly, the cells were collected after different treatments and washed thrice with cold phosphate‐buffered saline (PBS) by centrifugation for 5 min at 500 g and re‐suspended at a density of 1 × 106/ml. Cells (500 μl) were next stained with the Annexin V FITC (5 μl) and PI (10 μl) and placed in the dark for 15 min. The results were analyzed by flow cytometry (FACS CaliburTM, BD Biosciences). The data of fluorescence intensity were analyzed using the FlowJo software. Each experiment was repeated at least four times.
Annexin 5 fluorescein isothiocyanate fitc propidium iodide staining kit
Manufactured by Merck Group
Sourced in United States
Annexin V fluorescein isothiocyanate (FITC)‐propidium iodide (PI) staining kit is a laboratory tool used for the detection and analysis of apoptotic and necrotic cells. It combines the use of Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a nucleic acid dye, to differentiate between early apoptotic, late apoptotic, and necrotic cells.
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Lab products found in correlation
Facscalibur, by BD (1 mentions)
Mtt reagent, by Merck Group (1 mentions)
Hoechst 33342 solution, by Beyotime (1 mentions)
Beyoecl plus, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Sirtinol, by Merck Group (1 mentions)
B cell lymphoma 2 bcl 2, by Cell Signaling Technology (1 mentions)
Gapdh cat no 10494 1 ap, by Proteintech (1 mentions)
Bcl 2 associated x protein bax, by Cell Signaling Technology (1 mentions)
Radioimmunoprecipitation ripa lysis buffer, by Beyotime (1 mentions)
2 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide staining kit
1
Apoptosis Measurement by Annexin V-FITC/PI
2
Sirtinol-Induced Apoptosis Mechanisms
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SAL, Sirtinol and 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reagent were purchased from Sigma-Aldrich; Merck KGaA (Darmstadt, Germany). Hoechst 33342 solution, the BCA protein assay kit, radioimmunoprecipitation (RIPA) lysis buffer and BeyoECL Plus were purchased from Beyotime Institute of Biotechnology (Shanghai, China). The caspase-3 enzyme-linked immunosorbent assay (ELISA) kit was purchased from the Nanjing Jiancheng Bioengineering Research Institute (cat no. H076; Nanjing, China). The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining kit was obtained from Sigma-Aldrich; Merck KGaA. Antibodies against SIRT1 (cat no. 9475), phosphorylated (p)-FOXO3α (cat no. 9466), B-cell lymphoma 2 (Bcl-2; cat no. 3498) and Bcl-2 associated X protein (Bax; cat no. 2772) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). GAPDH (cat no. 10494-1-AP) and secondary antibodies were obtained from ProteinTech Group, Inc. (Chicago, IL, USA).
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