Cell apoptosis detection kit

Manufactured by Beyotime

Sourced in China

The cell apoptosis detection kit is a laboratory tool designed to detect and analyze the process of programmed cell death, known as apoptosis. The kit provides the necessary reagents and protocols to measure various markers associated with the apoptotic pathway, enabling researchers to study this fundamental biological phenomenon.

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Apoptosis detection kit (76 citations) Apoptosis kits (2 citations)

12 protocols using cell apoptosis detection kit

Assessing Viability and Apoptosis in mFB-MSC Co-cultures

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Viability was assayed using CellTiter-Blue^® cell viability assay kit and performed as per manufacturer’s instructions. For viability assay, MSCs in the co-culture were inactivated by 10 μg/mL mitomycin C (Dalian Meilun Biology Technology Co., Ltd, China) for 3 h before being mixed with mFBs. Cells were plated in 48-well plate with constant number of mFBs and an equal number of inactivated MSCs used in the co-culture among each ratio were plated in individual wells for background elimination. Before the assay, culture medium was discarded. Alamar blue solution prepared in fresh medium was added to each well and incubated at 37°C. Fluorescence values were read by microplate reader (Molecular Devices, USA) with excitation at 560 nm and emission at 590 nm. All values were normalized to the mFB mono cultures.
Cell apoptosis detection kit (Beyotime, China) was used to verify that the mFBs were viable in co-culture. All procedures were accomplished as per the instructions. Cells treated with 500 μmol/L H₂O₂ for 2 h were used as positive control.

Li X., Zhao H., Qi C., Zeng Y., Xu F, & Du Y. (2015). Direct intercellular communications dominate the interaction between adipose-derived MSCs and myofibroblasts against cardiac fibrosis. Protein & Cell, 6(10), 735-745.

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Apoptosis Detection by Flow Cytometry

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The apoptotic rate of cells was detected by flow cytometry using cell apoptosis detection kit (Beyotime, China). After plasmid transfection or cisplatin treatment, cells were digested, collected, and suspended with Annexin V-FITC binding buffer. Afterward, the cells were incubated with Annexin V-FITC and Propidium Iodide for 10 to 20 minutes in the dark room. Finally, the labeled cells were analyzed with a flow cytometer (Aceabio, USA).

Liu H., Song M., Sun X., Zhang X., Miao H, & Wang Y. (2021). T-box transcription factor TBX1, targeted by microRNA-6727-5p, inhibits cell growth and enhances cisplatin chemosensitivity of cervical cancer cells through AKT and MAPK pathways. Bioengineered, 12(1), 565-577.

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Cell Cycle and Apoptosis Measurement

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Cell cycle and cell apoptosis were both measured using FCM according to the reference protocol.30 (link) Briefly, cells were seeded into six-well plates to allow attachment. Then different formulations were added into each well and further incubated for 48 hours. The cells were then collected and further operations performed were according to the manufacturer’s instructions (cell cycle detection kit and cell apoptosis detection kit; Beyotime Biotechnology).

Shi M., Zhao X., Zhang J., Pan S., Yang C., Wei Y., Hu H., Qiao M., Chen D, & Zhao X. (2018). pH-responsive hybrid nanoparticle with enhanced dissociation characteristic for siRNA delivery. International Journal of Nanomedicine, 13, 6885-6902.

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Cell Cycle and Apoptosis Analysis

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Cell cycle and apoptosis were detected using a cell cycle kit and a cell apoptosis detection kit (Beyotime, Shanghai, China). The bovine preadipocytes cells were plated into 6-well plates and treated for 24 h. The cells were digested with trypsin, and centrifuged at about 1000× g for 3~5 min. Then, about 1 mL of pre-cooled PBS was added and the cells were resuspended. After re-centrifugation, the supernatant was discarded, and added into 1 mL of pre-cooled 70% ethanol, and gently blown and mixed. The cells were fixed at 4 °C for 24 h. After centrifugation at 1000× g for 3~5 min, the supernatant was discarded, and about 1 mL of pre-cooled PBS was added and the cells were resuspended. We added 0.5 mL of propidium iodide staining solution to each tube of cell samples, and the cell precipitate was slowly and fully re-suspended, incubated at 37 °C for 30 min in the dark, and detected by flow cytometry (BD-C6 Plus, Shanghai, China) (each treatment had three parallel replicates).

Hu C., Feng X., Ma Y., Wei D., Zhang L., Wang S, & Ma Y. (2023). CircADAMTS16 Inhibits Differentiation and Promotes Proliferation of Bovine Adipocytes by Targeting miR-10167-3p. Cells, 12(8), 1175.

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Apoptosis Measurement by Annexin V Staining

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Apoptosis was measured by Annexin V staining using a Cell Apoptosis Detection kit (Beyotime Biotechnology, Jiangsu, China) according to the manufacturer’s instructions, and the percentage of apoptotic cells was determined by flow cytometry (BDAccuri C6, BD Biosciences, Franklin Lakes, NJ, USA). Briefly, BGC823 stable cells infected with either the pLenti-shRNA-XIAP-AS1 or pLenti-shScramble cells were treated with 100 ng/ml TRAIL for 24 h, and the cells were then trypsinized and washed with cold PBS. The cells were harvested by centrifugation at 1,000 g for 5 min, and the pellet, containing approximately 1.0 × 10⁵ cells, was resuspended in 195 μl of binding buffer before being incubated with 5 μl of Annexin V-FITC and 10 μl of PI (Sigma-Aldrich, St Louis, USA) at room temperature for 10 min in the dark. The cells were subsequently collected and washed with cold PBS, and the percentage of apoptotic cells was analyzed using flow cytometry.

Cai J., Wang D., Bai Z.G., Yin J., Zhang J, & Zhang Z.T. (2017). The long noncoding RNA XIAP-AS1 promotes XIAP transcription by XIAP-AS1 interacting with Sp1 in gastric cancer cells. PLoS ONE, 12(8), e0182433.

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Apoptosis Detection in Decalcified Bone

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The bone was decalcified, paraffin embedded and section dewaxed to water as previously mentioned. The Cell Apoptosis Detection Kit (Beyotime, China) was used as per the manufacturer's instructions. Anti‐fluorescence quenching tablet encapsulation was utilized, and under a 400× high‐power fluorescence microscope, three fields were randomly selected in each section to observe the distribution of apoptotic cells.

Shi J., Zhang B., Wu Z., Zhang Y., Gupta A., Wang X., Wang J., Pan L., Xiao M., Zhang S, & Wang L. (2024). Peripheral nerve‐derived Sema3A promotes osteogenic differentiation of mesenchymal stem cells through the Wnt/β‐catenin/Nrp1 positive feedback loop. Journal of Cellular and Molecular Medicine, 28(8), e18201.

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Cytotoxicity of Nanoparticle Formulations

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Human PCA cell lines (PC-3, DU-145, and LNCap) were provided by Sure Bio-Tech Co., Ltd. (Shanghai, China). The cells were cultured in DMEM containing 10% fetal bovine serum (FBS) and 1% antibiotics. The medium, trypsin, and antibiotics were purchased from HyClone Co., Ltd. (UT, USA). The FBS was obtained from Gibco Co. (NY, USA). DOX, DOC, HA, coumarin-6, and IR-780 were purchased from Aladdin Corp. (Shanghai, China). The protein extraction kit, CCK-8 kit, TUNEL staining kit, and cell apoptosis detection kit were purchased from Beyotime Co., Ltd. (Shanghai, China). The antibodies (Bcl-2, Bax, Caspase 3, horseradish peroxidase-labeled secondary antibody) were supplied by Cell Signaling Co. (MA, USA). Other dyes and chemical reagents were obtained from Bokeri Co., Ltd. (Xi’an, China). The BALB/c mice and BALB/c-nu/nu mice were provided by Peking HFK Biotech Co., Ltd. (Beijing, China). CSaSt was synthesized in our lab.

Li K., Zhan W., Chen Y., Jha R.K, & Chen X. (2019). Docetaxel and Doxorubicin Codelivery by Nanocarriers for Synergistic Treatment of Prostate Cancer. Frontiers in Pharmacology, 10, 1436.

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Cell Apoptosis Detection by Flow Cytometry

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After centrifugation, cells were collected and stained with FITC Annexin V and propidium iodide using cell apoptosis detection kit (Beyotime, China) according to the manufacturer’s instructions. After rinsing, the signals of cells were analyzed by flow cytometry (BD Biosciences, USA) immediately.

Gong W, & Zhang S. (2023). YB1 participated in regulating mitochondrial activity through RNA replacement. Frontiers in Oncology, 13, 1145379.

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Cell Apoptosis Measurement Protocol

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The cells were digested with trypsin, and cells were centrifuged at 2000 g for 10 min. The cells were suspended with binding buffer containing 5 μL propidium iodide and 5 μL Annexin V-FITC with cell apoptosis detection kit (#C1062S, Beyotime, China). After incubation for 15 min in the dark, cell apoptosis was detected with Novocyte cytometer.

Xu Y., Chen Y., Yao M., You Y., Nie B., Zeng M, & Jiang H. (2023). MicroRNA-146a Improved Acute Lung Injury Induced by hepatic Ischemia-reperfusion Injury by Inhibiting PRDX1. Dose-Response, 21(2), 15593258231169805.

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Doxorubicin-Induced Apoptosis in U251 Cells

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The density of U251 cells was 1 × 10 5 cells per well (6-well plate). For apoptosis induction, cells were treated with 0, 100, 200,500 nM Doxorubicin (Selleck, USA) for 4 days after tests of several concentration gradients. Effects of apoptosis were evaluated using a Cell apoptosis detection kit (Beyotime, China). Apoptosis was induced in U251 cells successfully for three times with DMSO (Beyotime, China) as a vehicle control, and the cell samples were stored in Trizol reagent (Invitrogen, USA) for stepwise transcriptome RNA sequencing (RNA-seq).

Yu H., Yu J., Wang M, & Jiang X. (2024). Characterization of Prognostic Apoptosis-Related Gene Signature to Evaluate Glioma Immune Microenvironment and Experimental Verification. Genetic testing and molecular biomarkers, 28(1).

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