Livers were collected post 4 h intervention in different groups. Liver was fixed in 4% paraformaldehyde (PFA), sections (5 µm) were created and stained with hematoxylin and eosin according to standard procedures. Immunohistochemistry (IHC) was performed on paraffin sections as previously described (26 (link)). The TUNEL assay was performed by apoptosis detection kits (Beyotime Biotechnology, China) to detect apoptosis in liver tissues according to the manufacturer’s instructions. Images were captured by a Leica Upright Microscope DM2500, and calculated semi-quantitatively at magnification ×400 with Image Pro Plus software.
Apoptosis detection kit
Manufactured by Beyotime
Sourced in China, United States, United Kingdom
The Apoptosis detection kit is a laboratory tool designed to detect and analyze apoptosis, a form of programmed cell death, in cell samples. The kit provides the necessary reagents and protocols to identify and quantify apoptotic cells using established techniques.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Ros detection kit, by Beyotime (3 mentions)
Flow cytometry, by Beckman Coulter (3 mentions)
Cellquest software, by BD (3 mentions)
Flow cytometry, by BD (2 mentions)
Facscan flow cytometer, by BD (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Ros assay kit, by Beyotime (2 mentions)
Mitochondrial membrane potential detection kit, by Beyotime (2 mentions)
Cytoflex s, by Beckman Coulter (2 mentions)
Ldh cytotoxicity assay kit, by Abcam (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Curcumin, by Merck Group (2 mentions)
Dulbecco s modi ed eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Adenosine, by Merck Group (2 mentions)
Mtt t, by Abcam (2 mentions)
Gastrodin, by Merck Group (2 mentions)
Propyl gallate, by Merck Group (2 mentions)
Nih3t3 cells, by American Type Culture Collection (1 mentions)
76 protocols using apoptosis detection kit
1
Histological Analysis of Liver Apoptosis
2
Synthesis and Evaluation of Compound 9
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SN38 and valproic acid were purchased from Energy Chemical (Anhui Zesheng Technology Co., Ltd., Anqin, China). Other chemical reagents were obtained from Sinopharm Chemical Reagent Co., Ltd. (Sinopharm Group Co. Ltd., Shanghai, China). Compound 9 was dissolved in 0.05% DMSO. Thin-layer chromatography on 0.25 mm silicon gel plates (GF254) were purchased from Energy Chemical Co., Ltd. DEME culture medium was purchased from Hyclone. (Thermo Fisher Scientific, Logan, UT, USA). FBS was purchased from Gibco (Thermo Fisher Scientific, Logan, UT, USA). MTT, JC-1 probe, and apoptosis detection kits were purchased from Beyotime Biotechnology (Biyuntian Co., Ltd., Shanghai, China). Murine fibroblast NIH3T3 cells and human cervical cancer HeLa cells were purchased from ATCC of US. NMR spectra were recorded on Bruker DRX-300 NMR and Bruker DRX-400 NMR (Bruker Co., Bremen, Germany). Mass spectra (MS) were obtained using an Agilent LC-MS (ESI) (Agilent Co., Santa Clara CA, USA). Fluorescence imaging was observed on an Olympus IX71 fluorescence microscope (Olympus Co., Tokyo, Japan). Apoptosis was analyzed on CytoFLEX flow cytometry (Bechman Coulter, Pasadena, CA, USA).
3
Apoptosis Pathway Enzyme Analysis
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The activities of Caspase 3 and Caspase 9 in the liver samples were determined following the protocol for apoptosis detection kits (Beyotime, Shanghai, China, Catalog: C1116 and C1158). The protein concentration in the sample to be tested was detected by the Bradford method (Beyotime, Shanghai, China, P0006), and the enzyme activity units of Caspase 3 and Caspase 9 contained in a unit weight protein of the sample were calculated.
4
Characterization of H. pylori Strain G27
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H. pylori strain G27 (Courtesy of Prof. Bi Hongkai, Nanjing Medical University), calf serum, a Columbia blood agar base, a brain heart infusion (BHI,OXOID) medium, L-cysteine (L-cys) (AR 99%, MACKLIN), a fluorescence orthomicroscope (OLYMPUS, Tokyo, Japan), reactive oxygen species (ROS) detection kits (Beyotime), cell apoptosis 4’,6-diamidino-2-phenylindole (DAPI) detection kits (Beyotime), apoptosis detection kits (Beyotime), reverse transcription kits (Monad), reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) kits (Monad), a Lightcycler96 fluorescence ration PCR instrument (Roche, Germany), and a scanning electron microscope were used in the present study.
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H. pylori strain G27 (Courtesy of Prof. Bi Hongkai, Nanjing Medical University), calf serum, a Columbia blood agar base, a brain heart infusion (BHI,OXOID) medium, L-cysteine (L-cys) (AR 99%, MACKLIN), a fluorescence orthomicroscope (OLYMPUS, Tokyo, Japan), reactive oxygen species (ROS) detection kits (Beyotime), cell apoptosis 4’,6-diamidino-2-phenylindole (DAPI) detection kits (Beyotime), apoptosis detection kits (Beyotime), reverse transcription kits (Monad), reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) kits (Monad), a Lightcycler96 fluorescence ration PCR instrument (Roche, Germany), and a scanning electron microscope were used in the present study.
5
Apoptosis Analysis of TH-Z835 Treatment
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Cells (5 × 105 cells per well) were seeded in six-well plate overnight. Next, the cells were treated with either DMSO or TH-Z835 for 12 h or 24 h. For apoptosis analysis, cells were harvested by trypsinization and washed twice with ice-cold PBS, then stained with Annexin V-FITC and PI by Apoptosis Detection Kit (Beyotime, C1062) in the dark at room temperature for 10 min. Then cells were analyzed with the BD FACS AriaII and FlowJo software.
6
Cell Cycle and Apoptosis Detection
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For cell cycle detection, we digested and washed the cells with cold PBS and fixed them using 75% ethanol overnight. The fixed cells were centrifuged at 1,000 × g for 5 min at 4°C, and the supernatant was removed. The cells were resuspended using PI/RNase diluted in the staining buffer according to the protocol of the testing kit (Beyotime Biotechnology, Nantong, China). After incubation for 30 min from light, the cells were detected using the cytoFLEX Flow Cytometry System (Beckman-Coulter). As for apoptosis, the digested cells were washed with PBS and resuspended using binding buffer according to the apoptosis detection kit protocol (Beyotime Biotechnology, Nantong, China), after incubation with annexin-V and PI for 10 min from light, and the detection was performed using the Flow Cytometry System.
7
Apoptosis and Cell Cycle Analysis
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Transfected Hep 3B and Huh7 cells were processed with an Apoptosis Detection Kit (Beyotime, China), according to the manufacturer’s instructions. Briefly, transfected cells were washed twice with cold PBS. After incubation with 5 μL of FITC-Annexin V and 5 μL propidium iodide for 20 min in the dark, apoptosis of Hep 3B and Huh7 was detected using a flow cytometer (FACScan; BD Biosciences, USA).
For cell cycle analyses, cells were collected and incubated with 70% ethanol at 4°C overnight for fixation. The cells were washed twice with PBS and incubated with 100 μg/mL RNase A and 50 μg/mL propidium iodide for 1 h at 37°C. The percentage of cells in each phase of the cell cycle was then measured by flow cytometry (FACScan; BD Biosciences, USA).
For cell cycle analyses, cells were collected and incubated with 70% ethanol at 4°C overnight for fixation. The cells were washed twice with PBS and incubated with 100 μg/mL RNase A and 50 μg/mL propidium iodide for 1 h at 37°C. The percentage of cells in each phase of the cell cycle was then measured by flow cytometry (FACScan; BD Biosciences, USA).
8
Sensitizing CD90+ LCSCs to Magnetic Hyperthermia
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CD90+ LCSCs were incubated with the TMs coupled with anti-CD90 (CD90@TMs) (hyperthermia temperature, 43°C, called TMs group) for 24 hours and then were placed on an ACMF (f=200 kHz; I=20 A) to heat for 1 hour. Control group (cells incubated with DMEM) was not heated. Apoptosis rates were measured by FCM using an apoptosis detection kit (Beyotime Institute of Biotechnology, Shanghai, People’s Republic of China) according to the manufacturer’s instructions. The expression of HSP90 was determined by Western blotting after 10 minutes, 4 hours, 8 hours, 12 hours, and 24 hours of culture time after heat treatment. To determine if the inhibition of HSP90 could sensitize CD90+ LCSCs to magnetic hyperthermia, the expression of caspase-3 was examined by Western blotting 24 hours after heat treatment. To investigate the effect of HSP90 on the susceptibility of CD90+ LCSCs to hyperthermia susceptibility, 17-AAG was incubated with the cells before they were heated (called 17-AAG/TMs). The rate of apoptosis rates and the expression of HSP90 were then detected.
9
SCRIB Regulation of Apoptosis in Colorectal Cancer
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According to the manufacturer’s protocol, a one-step TUNEL (TdT-mediated dUTP gap end labelling) apoptosis detection kit (Beyotime, C1090) was used to examine the apoptosis of HCT116 and DLD-1 cells after the knockdown and overexpression of SCRIB, respectively. The cells were photographed under an Olympus FSX100 microscope (Tokyo, Japan).
10
Annexin V-FITC Apoptosis Assay in Endothelial Cells
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An apoptosis detection kit provided by Beyotime Biotechnology was used to detect and quantify apoptosis in vascular endothelial cells. Briefly, cells were trypsinised and resuspended at a concentration of 1 × 106/ml in diluted binding buffer and labelled with 10 μl of annexin V-FITC. Cells were incubated for 30 min at room temperature in dark, followed by a 5-min incubation with 5 μl of PI. Subsequently, 400 μl of 1× binding buffer was added to each tube. Flow cytometric analysis was performed to monitor the annexin V and DNA-bound PI. Data acquisition and analysis were performed using the FlowJo software.
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