Chlorin E6, Atovaquone, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC HCl), and N-carboxythiosuccinimide (sulfo-NHS) were purchased from Shanghai Yuanye Biotechnology Co., Ltd. Dimethyl sulfoxide (DMSO) was purchased from Beijing LABGIC technology Co., Ltd; monodisperse carboxylic magnetic mesoporous silica nanomaterials (Fe3O4@MSNs) were purchased from Nanjing NANOEAST Biotech Co., Ltd; targeted peptide (iRGD) was purchased from Hefei Bank Peptide Biological technology Co., Ltd; 1,3-diphenylisobenzofuran (DPBF) was purchased from Shanghai Macklin Biochemical Co., Ltd; PBS buffer solution was purchased from Beijing Rangeke Technology Co., Ltd; DMEM medium, peptide bovine serum (FBS), trypsin, EDTA-free trypsin and double antibody (PS) were purchased from Thermo Fisher Scientific. 4′,6-Diamidino-2-phenylindole (DAPI), 4% paraformaldehyde, and electron microscope fixative were purchased from Wuhan Servicebio Technology Co., Ltd; cck8 kit and AnnexinV-FITC/PI double staining apoptosis detection kit were purchased from Jiangsu KeyGEN BioTECH Co., Ltd; 2′,7′-dichlorofluorescin diacetate (DCFH-DA) and calcein/PI cell activity and cytotoxicity detection kits were purchased from Shanghai Beyotime Biotechnology Co., Ltd.
Annexin 5 fitc pi double staining apoptosis detection kit
Manufactured by Keygen Biotech
Sourced in China
The Annexin V-FITC/PI double staining apoptosis detection kit is a laboratory tool designed to detect and quantify apoptosis in cell samples. It utilizes the binding properties of Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a DNA-binding dye, to identify cells undergoing apoptosis. The kit provides a reliable method for distinguishing between viable, early apoptotic, late apoptotic, and necrotic cells through fluorescence-based analysis.
Automatically generated - may contain errors
Lab products found in correlation
Cell cycle detection kit, by Keygen Biotech (2 mentions)
Electron microscope fixative, by Wuhan Servicebio Technology (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Edta free trypsin, by Thermo Fisher Scientific (1 mentions)
2 7 dichlorofluorescin diacetate dcfh da, by Beyotime (1 mentions)
Cck 8 kit, by Keygen Biotech (1 mentions)
Rnase a, by Illumina (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Accuri c6, by BD (1 mentions)
Novocyte, by Agilent Technologies (1 mentions)
Facs flow cytometer, by Thermo Fisher Scientific (1 mentions)
Annexin 5, by Keygen Biotech (1 mentions)
6 protocols using annexin 5 fitc pi double staining apoptosis detection kit
1
Multifunctional Nanomaterial-based Theranostics
2
Apoptosis and Cell Cycle Analysis
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Apoptosis was detected using an Annexin V-FITC/PI double-staining apoptosis detection kit (KeyGen Biotech, KGA107) according to manufacturer instructions. Cells were collected after treatment with trypsin without EDTA (Solarbio, T1350) and washed once with PBS. The cell pellet was resuspended in binding buffer, and 5 μL Annexin V-FITC and PI staining agents were added and incubated for 15 and 5 min in the dark, respectively. Then, the apoptosis rate of each group was determined with data obtained with a CytoFLEX Flow Cytometer (Beckman Coulter).
For cell-cycle analysis, cells were fixed in 70% ice-cold ethanol overnight. Next, 30 μL RNase A (Epicenter, RNR07250) was incubated with the cells at 37°C for 30 min. The cells were then stained at 4°C for 30 min with 120 μL PI (KeyGen Biotech, KGA512) in the dark. Fluorescence intensity was measured using a CytoFLEX Flow Cytometer (Beckman Coulter). The proportion of cells in the G0/G1, S, and G2/M cell-cycle phases was determined, and the proportions were then compared.
For cell-cycle analysis, cells were fixed in 70% ice-cold ethanol overnight. Next, 30 μL RNase A (Epicenter, RNR07250) was incubated with the cells at 37°C for 30 min. The cells were then stained at 4°C for 30 min with 120 μL PI (KeyGen Biotech, KGA512) in the dark. Fluorescence intensity was measured using a CytoFLEX Flow Cytometer (Beckman Coulter). The proportion of cells in the G0/G1, S, and G2/M cell-cycle phases was determined, and the proportions were then compared.
3
Cell Apoptosis and Cycle Assays
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MCE provided PA (CAT: HY-N0371, United States). Punosai Co., Ltd. provided HGC-27 (CAT: CL-0107, China) and GES-1 (CAT: CL-0563, China) cells, which were confirmed by STR to be free of cross-contamination. Gemini was used to sell fetal bovine serum (CAT: 900108, China). Gibco provided RPMI-1640 medium (CAT: 1187517, United States). Dojindo Cell Counting Kit-8 (CAT: CK04, Japan). The Annexin-V FITC/PI double-staining apoptosis detection kit (CAT: KGA1023; China) and cell cycle detection kit (CAT: KGA511, China) were provided by Jiangsu KeyGEN Bio TECH Corp., Ltd. Antibodies against PI3K (CAT: ab133595, United Kingdom), p-PI3K (CAT: ab278545, United Kingdom), BAX (CAT: ab32503, United Kingdom), BCL-2 (CAT: ab182858, United Kingdom), Caspase 3 (CAT: ab32351, United Kingdom), Caspase 9 (CAT: ab32539, United Kingdom), and GADPH (CAT: ab8245, United States) were purchased from Abcam. Antibodies against AKT (CAT: AF6261, China) and p-AKT (CAT: AF0016, China) were purchased from Affinity.
4
Cell Cycle and Apoptosis Analysis in HL60 Cells
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Cell cycle analysis using Cell Cycle Detection Kit (KeyGEN, Cat# KGA512) was performed with Ara-C, Aprepitant or the combination of both drugs at 37°C for 24 hours. Then, HL60 cells were collected and were washed with PBS two times before fixing with methanol at −20°C overnight. Cells were stained with the mixed solution of PI and RNase A and then incubated at room temperature for 30 min prior to flow cytometry analysis (BD Accuri™ C6). NovoExpress (aceabio, USA) software was used for analysis results.
Assessment of cell death was detected by Annexin V-FITC/PI double staining Apoptosis Detection Kit (KeyGEN, Cat# KGA108). HL60 cells were treated with Ara-C, Aprepitant or the combination of both drugs for 24 hours. Cells were harvested and washed with PBS two times, and then resuspended in 500 μL of binding buffer containing 5 μL Annexin V-FITC and 5 μL propidium iodide (PI) for 15 min at room temperature, followed by Flow Cytometry (BD Accuri™ C6). NovoExpress (aceabio, USA) software was used for analysis results.
Assessment of cell death was detected by Annexin V-FITC/PI double staining Apoptosis Detection Kit (KeyGEN, Cat# KGA108). HL60 cells were treated with Ara-C, Aprepitant or the combination of both drugs for 24 hours. Cells were harvested and washed with PBS two times, and then resuspended in 500 μL of binding buffer containing 5 μL Annexin V-FITC and 5 μL propidium iodide (PI) for 15 min at room temperature, followed by Flow Cytometry (BD Accuri™ C6). NovoExpress (aceabio, USA) software was used for analysis results.
5
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis in BC cells was measured by Annexin V-FITC/PI double staining apoptosis detection kit (KGA1102, KeyGEN BioTech, China). After centrifugation, cells were resuspended in 500 μL binding buffer. Then, 5 μL AnnexinV-FITC and 5 μL PI were separately added and mixed. Cells were incubated for 15 min in the dark, followed by flow cytometry detection (NovoCyte, Agilent, USA).
6
Apoptosis Assay of Transfected Cells
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The transfected TPC-1 and CAL62 were detached by trypsinization and then resuspended in the binding buffer of annexin V-FITC/PI double staining apoptosis detection kit (KeyGEN BioTech, Jiangning, Nanjing, China). Cells were then stained with 5 µL of PI and 5 µL of annexin V (KeyGEN BioTech). The apoptotic ratio was analyzed with a FACS flow cytometer (Life Technologies, Darmstadt, Germany).
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