Cells were seeded at a density of 1 × 105 cells/well in a 12-well plate. At the 24-h point, cell morphology was assessed using the Apoptosis and Necrosis Assay Kit (Beyotime, Shanghai, China, Cat No. C0003). The cells were stained with Hoechst 33342 and propidium iodide (PI) according to the manufacturer’s protocols and then visualized by fluorescence microscopy. An Annexin-V-FITC/PI Cell Apoptosis Detection Kit (Meilunbio, Dalian, China, Cat No. MA0220-2) was used to detect the cells’ apoptotic rate according to the manufacturer’s direction. Afterward, the apoptotic rate of cells was measured within 1 h by a flow cytometer (BD Biosciences).
Apoptosis and necrosis assay kit
Manufactured by Beyotime
Sourced in China
The Apoptosis and Necrosis Assay Kit is a laboratory equipment product that enables the detection and quantification of apoptosis and necrosis in cells. The kit provides the necessary reagents and protocols to perform these analyses.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700, by Zeiss (2 mentions)
Lsm 510, by Zeiss (2 mentions)
Annexin 5 fitc pi cell apoptosis detection kit, by Meilun (1 mentions)
Operetta cls high content cell imaging analysis system, by PerkinElmer (1 mentions)
Secondary antibody dilution buffer, by Beyotime (1 mentions)
Primary antibody dilution buffer, by Beyotime (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Sds page gel quick preparation kit, by Beyotime (1 mentions)
Cell lysis buffer for western and ip, by Beyotime (1 mentions)
Polyvinylidene fluoride pvdf membrane, by Beyotime (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Blocking buffer, by Beyotime (1 mentions)
Beyoecl plus, by Beyotime (1 mentions)
Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Z vad fmk, by Selleck Chemicals (1 mentions)
Cell cycle and apoptosis analysis kit, by Beyotime (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by Beyotime (1 mentions)
Hoechst 33342 pi, by Beyotime (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
4 phenylbutyrate 4 pba, by Merck Group (1 mentions)
46 protocols using apoptosis and necrosis assay kit
1
Apoptosis and Necrosis Assessment
2
Apoptosis and Necrosis Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Nuclear condensation, and the apoptotic or necrotic cells were evaluated by Apoptosis and Necrosis Assay kit (Beyotime, China) (Song et al., 2011 (link)). In brief, the treated cells were harvested, washed, and stained with 5 μg/ml Hoechst33342 and 5 μg/ml propidium iodide (PI) for 20–30 min at 4°C. After that, the cells were collected, washed once, and examined using a Nikon Intensilight C-HGFI microscope (Nikon, Japan).
3
Anticancer Effects of Re-CDs Assessed
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Logarithmic growth phase cancer cells were treated with different concentrations of Re-CDs in 96-well plates for 24 hours. Then, high-content cell imaging analysis was undertaken to visually reflect the anticancer effect of Re-CDs. An apoptosis and necrosis assay kit (Beyotime Biotechnology) was used in the experiment and the staining assay was carried out as follows. Cells were seeded in 96-well plate and treated with different concentrations of Re-CDs. After 24 hours of incubation, the supernatant was removed and cells were washed with cold PBS. Cancer cells were labeled with Hoechst 33342 and propidium iodide (PI) for 30 minutes at 4°C under working concentration. Cells were washed with PBS three times to remove redundant dye liquors. Images were obtained with the Operetta CLS™ high-content cell imaging analysis system (PerkinElmer Inc., Waltham, MA, USA). The 20× objective lens was applied in each treatment condition. Results were represented by over 40 fields per plate, leading to fields overall acquired for each condition. As we tried to investigate the effect of ROS on apoptosis, the antioxidant N-acetyl-l -cysteine (NAC)- and Re-CDs-treated groups were taken as comparison.
4
Quercetin and Rutin Cytotoxicity Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quercetin, rutin, dimethyl sulfoxide (DMSO) and 5-fluorouracil were purchased from Shanghai Kaiyang Biotechnology Company (Shanghai, China). Acetonitrile (HPLC), RPMI Medium 1640, and fetal bovine serum (FBS) were products of Gibco Life Technologies (NY, USA). The Apoptosis and Necrosis Assay Kit was purchased from the Beyotime; OLYMPUS IX73. Guava easyGyte; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), Cell lysis buffer for Western and IP, the SDS-PAGE Gel Quick Preparation Kit, Protein Marker, Polyvinylidene Fluoride (PVDF) Membrane, Blocking Buffer, Primary Antibody Dilution Buffer, Secondary Antibody Dilution Buffer, and BeyoECL Plus were purchased from the Beyotime Institute of Biotechnology (Shanghai, China).
5
Synthesis and Evaluation of Acridine Derivative
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
N-(2-(dimethylamino)ethyl)-1-((3-methoxybenzyl)amino)-5-nitro-9-oxo-9,10-dihydro-acridine-4-carboxamide (8q ) was synthesized by Zhang [4 (link)]. Z-VAD-FMK (a pan-caspase inhibitor) was purchased from Selleck (Shanghai, China). Human chronic myelogenous leukemia K562 cells were purchased from the Chinese Academy of Sciences Cell Bank. Human chronic myelogenous leukemia KCL-22 cells and K562 adriamycin-resistant cells (K562/ADR) were purchased from Zhen Shanghai and Shanghai Industrial Co., Ltd. (China). Iscove’s Modified Dubecco’s Medium (IMDM) and Fetal bovine serum (FBS) was purchased from Hyclone (Logan, Utah, USA). Cleaved Caspase-3 antibodies were purchased from Cell Signaling Technology, Inc. (Boston, Massachusetts, USA), Cleaved Caspase-9, 8 antibodies were purchased from Wuhan SanYing Co., Ltd. (Wuhan, China); other antibodies we used in this study were purchased from Beyotime Biotechnology (Shanghai, China). Annexin V-FITC/PI apoptosis detection kit, apoptosis and necrosis assay kit and cell cycle and apoptosis analysis kit were purchased from Beyotime Biotechnology (Shanghai, China). Formic acid (HPLC) was purchased from Tedia (Ohio, USA). Acetonitrile (HPLC) and methanol (HPLC) were purchased from Fisher (Waltham, Massachusetts, USA).
6
Apoptosis and Necrosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hoechst 33342/PI (propidine iodide) apoptosis and Necrosis Assay Kit was purchased from Beyotime (China, Catalogue No. C1056). Washed cells twice with cold PBS and added 5 μl of Hoechst 33342 staining solution. Mixed gently, incubated at 4 °C for 20–30 min in an ice bath. Then observed with a microscope ant took photos.
7
Hoechst 33342/PI Staining Assay for Apoptosis and Necrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hoechst 33342/PI staining assay was performed according to the manufacturer's instructions (Apoptosis and Necrosis Assay Kit, Beyotime Institute of Biotechnology). Briefly, HepG2 and Hepa1-6 cells were cultured in 35 mm cover glass-bottom culture dishes (Corning Incorporated, Corning, NY, USA) and subjected to the indicated treatments. The final concentration of DMSO was less than 0.1%, and DMSO (0.1%) served as vehicle control. Then, Hoechst 33342 (5 μL) and PI dye (5 μL) were added and then incubated for 20 min at 4 °C in the dark and visualized under a confocal scanning microscope (Zeiss LSM 700, Jena, Germany).
8
Macrophage Apoptosis and Necrosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse peritoneal macrophages were incubated with DMSO or Munronoid I (50 μM) for 2 h, then stimulated with LPS (100 ng/ml) for 4 h. The cells were treated with ATP (5 mM) for 1h, then the cells were washed twice with cold PBS and incubated with Hochest33342 (5 μg/ml; staining for all cells) and propidium iodide (PI, 2 μg/ml; staining for membrane-damaged cells) double staining of Apoptosis and Necrosis assay Kit (Beyotime Biotechnology, Shanghai, China) at 4°C for 20 min. After that, the treated cells were washed once by ice-cold PBS immediately and observed under fluorescence confocal microscopy.
9
Apoptosis and Endoplasmic Reticulum Stress Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ZEA and 4-phenylbutyrate (4-PBA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). An Annexin V-PE/7-AAD kit, Total Protein Extraction Kit and BCA Protein Assay Kit were purchased from Nanjing Keygen Biotech Co., Ltd. (Nanjing, Jiangsu, China). The Apoptosis and Necrosis Assay Kit was purchased from Beyotime Institute of Biotechnology (Shanghai, China). Anti-β-actin antibody was obtained from Beijing CWBIO Co., Ltd. (Beijing, China). Anti-GRP78 and anti-CHOP antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
10
Apoptosis and Necrosis Assay in Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the apoptosis or necrosis phenomenon, Hoechst 33342 and PI staining assays were carried out. For these, 1 × 105 RAW 264.7 macrophages per well were cultured over night in 24-well plates and treated with 0, 30 and 50 μM ZEA for 12 and 24 h. The resulting cell morphology was assessed using an Apoptosis and Necrosis Assay Kit (Beyotime Institute of Biotechnology, Shanghai, China). Briefly, cells in 24-well plates were trypsinized and washed twice with PBS. The cells were then stained with Hoechst 33342 and PI for 30 min at 4 °C in the dark. Apoptotic and necrotic cells were observed using a Nikon epifluorescence microscope (Nikon Eclipse 80i; Nikon, Tokyo, Japan).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!