N,N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO) and doxorubicin (DOX) were provided from Shanghai Macklin Biochemical Co., Ltd. Benzoic acid (BA) and zirconium chloride octahydrate (ZrOCl2·8H2O, 99.99%) were provided by Aladdin Biochemical Technology Co., Ltd. Tetrakis(4-carboxyphenyl)porphyrin (TCPP) were obtained from Beijing Innochem Technology Co., Ltd. 1,3-Diphenylisobenzofuran (DPBF) were acquired from Adamas Reagent Co., Ltd. Penicillin/streptomycin, Dulbecco's phosphate buffered saline (PBS), [4′5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium-bromide (MTT), and trypsin were provided by Shanghai Sigma Aldrich Trading Co., Ltd. 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA), Hoechst 33342, Annexin V-FITC/PI cell apoptosis kit and Calcein-AM/PI double staining kit were provided by Shanghai Beyotime Biotechnology Co., Ltd. Fetal bovine serum (FBS) was obtained from Gibco reagent Co., Ltd. Roswell Park Memorial Institute 1640 (RPMI-1640) medium and Dulbecco's modified Eagle's medium (DMEM) were purchased from Biosharp reagent Co., Ltd. All chemicals were of analytical grade and were not purified before use. Nucleic acid sequences were synthesized and HPLC purified by Shanghai Sangon Biotech Co., Ltd as Table S1.
Annexin 5 fitc pi cell apoptosis kit
Manufactured by Beyotime
Sourced in China
The Annexin V-FITC/PI cell apoptosis kit is a laboratory reagent used to detect and quantify apoptosis, a process of programmed cell death. The kit contains Annexin V-FITC, a fluorescent conjugate that binds to phosphatidylserine residues exposed on the cell surface during apoptosis, and propidium iodide (PI), a DNA-binding dye that stains cells with compromised cell membranes. The kit allows for the simultaneous detection and differentiation of early apoptotic, late apoptotic, and necrotic cell populations using flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Fluorescence activated cell sorting (facs), by BD (2 mentions)
Cell cycle staining kit, by MultiSciences Biotech (2 mentions)
Calcein am pi double staining kit, by Beyotime (1 mentions)
Doxorubicin dox, by Maclin Biochemical Technology (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
2 7 dichlorodihydrofluorescein diacetate dcfh da, by Beyotime (1 mentions)
Attune nxt flow, by Thermo Fisher Scientific (1 mentions)
Lsrfortessa, by BD (1 mentions)
Calcein, by Beyotime (1 mentions)
Benzoic acid, by Sinopharm (1 mentions)
Field emission scanning electron microscope, by Merck Group (1 mentions)
Zetasizer nano zs zen3600, by Malvern Panalytical (1 mentions)
Miniflex 600 x ray diffractometer, by Rigaku (1 mentions)
2 7 dichlorofluorescin diacetate dcfh da, by Beyotime (1 mentions)
Thiazolyl blue tetrazolium bromide, by Merck Group (1 mentions)
Alloxan, by Merck Group (1 mentions)
Pancreatin, by Merck Group (1 mentions)
Normal saline, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
8 protocols using annexin 5 fitc pi cell apoptosis kit
1
Multifunctional Nanoparticle Platform for Cancer Therapy
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N,N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO) and doxorubicin (DOX) were provided from Shanghai Macklin Biochemical Co., Ltd. Benzoic acid (BA) and zirconium chloride octahydrate (ZrOCl2·8H2O, 99.99%) were provided by Aladdin Biochemical Technology Co., Ltd. Tetrakis(4-carboxyphenyl)porphyrin (TCPP) were obtained from Beijing Innochem Technology Co., Ltd. 1,3-Diphenylisobenzofuran (DPBF) were acquired from Adamas Reagent Co., Ltd. Penicillin/streptomycin, Dulbecco's phosphate buffered saline (PBS), [4′5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium-bromide (MTT), and trypsin were provided by Shanghai Sigma Aldrich Trading Co., Ltd. 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA), Hoechst 33342, Annexin V-FITC/PI cell apoptosis kit and Calcein-AM/PI double staining kit were provided by Shanghai Beyotime Biotechnology Co., Ltd. Fetal bovine serum (FBS) was obtained from Gibco reagent Co., Ltd. Roswell Park Memorial Institute 1640 (RPMI-1640) medium and Dulbecco's modified Eagle's medium (DMEM) were purchased from Biosharp reagent Co., Ltd. All chemicals were of analytical grade and were not purified before use. Nucleic acid sequences were synthesized and HPLC purified by Shanghai Sangon Biotech Co., Ltd as Table S1.†
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N,N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO) and doxorubicin (DOX) were provided from Shanghai Macklin Biochemical Co., Ltd. Benzoic acid (BA) and zirconium chloride octahydrate (ZrOCl2·8H2O, 99.99%) were provided by Aladdin Biochemical Technology Co., Ltd. Tetrakis(4-carboxyphenyl)porphyrin (TCPP) were obtained from Beijing Innochem Technology Co., Ltd. 1,3-Diphenylisobenzofuran (DPBF) were acquired from Adamas Reagent Co., Ltd. Penicillin/streptomycin, Dulbecco's phosphate buffered saline (PBS), [4′5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium-bromide (MTT), and trypsin were provided by Shanghai Sigma Aldrich Trading Co., Ltd. 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA), Hoechst 33342, Annexin V-FITC/PI cell apoptosis kit and Calcein-AM/PI double staining kit were provided by Shanghai Beyotime Biotechnology Co., Ltd. Fetal bovine serum (FBS) was obtained from Gibco reagent Co., Ltd. Roswell Park Memorial Institute 1640 (RPMI-1640) medium and Dulbecco's modified Eagle's medium (DMEM) were purchased from Biosharp reagent Co., Ltd. All chemicals were of analytical grade and were not purified before use. Nucleic acid sequences were synthesized and HPLC purified by Shanghai Sangon Biotech Co., Ltd as Table S1.
2
Single-Cell Suspension Preparation and Apoptosis Analysis
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The preparation of single‐cell suspension was as follows. After the rats in each group were sacrificed, the brain tissues were quickly separated on ice. About 30 mm3 brain tissues around the hematoma were cut using an ophthalmic scissor and then cut into pieces in a culture dish containing 1 ml of PBS. The tissues were added 50 μl of 0.25% trypsin, mashed several times using a tube, and then filtered through a 200‐mesh filter into a centrifuge. PBS was added to dilute to 5 ml. Next, and the centrifugation was carried out at 1,000 rpm for 15 min. After that, the tissues were rinsed twice with PBS, fixed with ethanol for 3 hr. The number of cells was 1 × 106/ml. The detection of cell apoptosis was performed using Attune NxT flow cytometry (ThermoFisher), and in accordance with the operation instructions of Annexin V‐FITC/PI cell apoptosis kit (Beyotime).
3
Cell Cycle and Apoptosis Analysis
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Cell cycle and apoptosis assays were analyzed by flow cytometry analysis using FACS (BD Biosciences). For the cell cycle analysis, cells were harvested by trypsinization and then fixed in ice-cold 75% ethanol at -20°C overnight. The next day, the cells were stained with propidium iodide (PI) according to the Cell Cycle Staining Kit (MultiSciences, China, CCS012). Cell cycle distributions were determined by ModFitLT Software. For the cell apoptosis assay, cells were carefully harvested by trypsinization and stained using an Annexin V-FITC/PI Cell Apoptosis Kit (Beyotime, C1062). The percentage of apoptotic cells was analyzed by FlowJo software.
4
Apoptosis Quantification by Flow Cytometry
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Cell apoptosis was measured using the Annexin-V-FITC/PI cell apoptosis kit (Beyotime Institute of Biotechnology). After treatment with 0.25% trypsin at 37°C for 2 min, 5×106 cells were collected, suspended in binding buffer and stained with 5 µl Annexin-V-FITC at 4°C for 15 min in the dark. Then, 5 µl PI was added to cells and incubated at 4°C for 5 min. Finally, cell apoptosis rate was measured using LSRFortessa™ (BD Biosciences) and analyzed with FlowJo 10.7 software (BD Biosciences). The cells with the Annexin-FITC label were regarded as apoptotic cells.
5
Multicomponent Luminescent Hybrid Materials
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Yttrium oxide (Y2O3), Ytterbium oxide (Yb2O3), Erbium oxide (Er2O3), and Polyacrylic acid polymers (PAA) were taken by Aladdin-Reagent Co. Ltd. (Shanghai, China). Zirconyl chloride octahydrate (ZrOCl2·8H2O) and benzoic acid (BA) were purchased from Sinopharm Chemical Reagent Co., Ltd., (Shanghai, China). Tetrakis (4-carboxyphenyl) porphyrin (TCPP) was synthesized based on the previous procedure with some modifications [1 (link)]. 2′,7′-dichlorofluorescin diacetate (DCFH-DA), Annexin V-FITC/PI Cell Apoptosis Kit, and Calcein were obtained from Beyotime Institute of Biotechnology (Shanghai, China) ScalaSansLF-Regular. All cell lines (SCC-7 and COS-7) were purchased from China Centre for Type Culture Collection (CCTCC). TEM images were carried out using Tecnai G20 S-TWIN operated at 200 kV. SEM images were taken by a field emission scanning electron microscope (Sigma). PXRD analysis was tested by Rigaku MiniFlex 600 X-ray diffractometer with Cu (Kα = 1.5418 Å). The hydrodynamic size and zeta potential were performed by dynamic light scattering (DLS) on a Malvern Zetasizer Nano-ZS ZEN3600 instrument.
6
In Vitro Evaluation of Antioxidant and Cytoprotective Effects
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CAP was provided by Shanghai Fengni Pharmaceutical Technology Co., Ltd. (Shanghai, China). DON was purchased from Sigma-Aldrich (St. Louis, MO, USA), dissolved in dimethylsulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA), formulated into 1 mg/mL stock solution, and stored at -20°C. H2O2, phosphate-buffered saline (PBS, pH = 7.4), 0.25% pancreatin with or without ethylenediaminetetraacetic acid (EDTA), penicillin-streptomycin, thiazolyl blue tetrazolium bromide (MTT), alloxan, and normal saline were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco's Modified Eagle Medium (DMEM), high-glucose DMEM, and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). The reactive oxygen species (ROS) and Annexin V-FITC/PI cell apoptosis kit were purchased from Beyotime Bioengineering Institute (Shanghai, China). Glibenclamide was purchased from Meryer Chemical Technology Co., Ltd. (Shanghai, China). Liuweidihuang pill was purchased from Beijing Tongrentang Co., Ltd. (Beijing, China).
7
Cell Cycle and Apoptosis Analysis
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Cell cycle and apoptosis assays were analyzed by flow cytometry analysis using FACS (BD Biosciences). For the cell cycle analysis, cells were harvested by trypsinization and then fixed in ice-cold 75% ethanol at − 20 °C overnight. The next day, the cells were stained with propidium iodide (PI) according to the Cell Cycle Staining Kit (MultiSciences, China, CCS012). Cell cycle distributions were determined by ModFitLT Software. For the cell apoptosis assay, cells were carefully harvested by trypsinization and stained using an Annexin V-FITC/PI Cell Apoptosis Kit (Beyotime, C1062). The percentage of apoptotic cells was analyzed by FlowJo software.
8
Annexin V-FITC/PI Apoptosis Assay
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For apoptosis detection, 1×105 HPAECs were treated with Annexin V (5 μL) and propidium iodide (PI) (5 μL) together for 15 minutes at room temperature in the dark. After cells were collected and washed with cold PBS twice, cell apoptosis was detected using an Annexin V-FITC/PI cell apoptosis kit (C1062L, Beyotime, China). Cell apoptosis was detected with Guava easyCyte Benchtop Flow Cytometer (BR168323; Luminex, Austin, TX) and data were analyzed using Kaluza C Analysis Software (version 2.1, Beckman Coulter, Indianapolis, IN).
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