Apoptosis detection kit

Manufactured by BestBio

Sourced in China

The Apoptosis detection kit is a laboratory tool used to detect and analyze programmed cell death, also known as apoptosis. The kit provides reagents and protocols to identify and quantify apoptotic cells in various sample types.

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Lab products found in correlation

Pi rnase staining solution, by BD (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Propidium iodide, by Beyotime (1 mentions) 8 mm pore transwell inserts, by Corning (1 mentions) Coulter dna prep reagents kit, by Beckman Coulter (1 mentions) Tgf β1, by Corning (1 mentions) Tgf β1, by Beyotime (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Tgf β1, by Thermo Fisher Scientific (1 mentions) Matrigel, by BD (1 mentions) Cell counting kit 8 cck 8 assay, by Beyotime (1 mentions) Calibur flow cytometer, by BD (1 mentions) Cell cycle detection kit, by BestBio (1 mentions) Facscalibur, by BD (1 mentions) Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions) Trisodium citrate dehydrate, by Merck Group (1 mentions) 1 ethyl 3 3 dimethylaminopropyl carbodiimide edc, by Merck Group (1 mentions) N hydroxysuccinimide nhs, by Merck Group (1 mentions) Cellquest flow cytometry software, by BD (1 mentions)

11 protocols using apoptosis detection kit

Cell Apoptosis and Cell Cycle Analysis

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Single cell suspensions were collected and washed twice with cold PBS. For apoptosis assays, AGS and MKN45 cells were double stained with propidium iodide (PI; Beyotime, Shanghai, China) and Annexin V-fluorescein isothiocyanate (FITC) by an apoptosis detection kit (BestBio, Shanghai, China).
For the cell cycle assay, the cells were fixed with cold 75% ethanol and stored at −20°C for at least 24 h. Then, cells were washed twice with cold PBS before 1 × 10⁶ cells were resuspended in 1 mL PI/RNase staining solution (BD Biosciences, USA) and finally analyzed by a FACSCalibur flow cytometer (BD Biosciences, USA).

Xie N., Pan Y., Wu J., Bai Y., Xiao C., Gao X., Wang J, & Liu N. (2021). MicroRNA-302s Might Regulate ARL4C-Mediated Gastric Cancer Progression via p53 Signaling: Bioinformatics Analysis and Experiments Validation. OncoTargets and therapy, 14, 2541-2553.

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Profiling TGFβ1-induced Senescence in AML Cells

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AML cell lines KG-1a was ordered from the Shanghai cell bank of the Chinese Academy of Sciences and cultured in RPMI 1640 (Gibco) supplemented with 20% fetal bovine serum. Cells were maintained at 37 °C in a 5% CO₂ incubator. KG-1a was precultured with 10 ng/ml TGFβ1 (PeproTech) for 48 h and used for Cell Counting Kit-8 (CCK8) assay (Beyotime Institute of Biotechnology, China), apoptosis detection (Apoptosis Detection Kit, BestBio), cell cycle detection (COULTER DNA PREP Reagents Kit, Beckman). Cell cycle phase distributions were generated by fitting DNA content histograms to applicable models using ModFit LT software for Windows (Version 5.0). KG-1a cells were stained with Senescence‐associated‐β‐Galactosidase (SA‐β‐Gal) staining (Beyotime) according to the instructions after TGFβ1 treatment for 7 days, and cells that stained green‐blue were evaluated as positive senescent cells. Cell suspension (1 × 10⁵, 200 μl of serum-free medium) after TGFβ1 pretreatment was seeded onto 8-mm Pore Transwell Inserts (Corning) in 24-well plates with or without matrigel (BD biosciences) and photographed within 24 h.

Liang X., Li C., Fan M., Zhang W., Liu L., Zhou J., Hu L, & Zhai Z. (2023). Immune-related lncRNAs pairs prognostic score model for prediction of survival in acute myeloid leukemia patients. Clinical and Experimental Medicine, 23(8), 4527-4538.

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Apoptosis Analysis of TMZ-Treated Cells

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Cells were dispensed in 6-well plates and treated with TMZ for 3 days. The cells were then collected and fixed with 70% ethanol for 24 h at 4°C, and the cell cycle was analyzed by Beijing Dingguo Changsheng Biotechnology Co. Ltd. (Beijing, China). The cells were collected and resuspended in 400 μL of ice-cold PBS for apoptosis analysis. An apoptosis detection kit (BestBio, Shanghai, China) was used to stain the cells, and Annexin-V FITC and 7-AAD were detected using a BD Calibur flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA).

Zhou J., Tong F., Zhao J., Cui X., Wang Y., Wang G., Kang C., Liu X, & Wang Q. (2023). Identification of the E2F1-RAD51AP1 axis as a key factor in MGMT-methylated GBM TMZ resistance. Cancer Biology & Medicine, 20(5), 385-400.

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Apoptosis Assay of Transfected Cells

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The HT29 and SW620 cells were seeded in a 6-well plate after being transfected with an interfering or overexpressing vector for 48 h. Then, the cells were digested with trypsin and 1 × 10⁶ cells were counted for analysis. The cells were washed with cold PBS three times. They were then collected and suspended into a single cell in a binding buffer. According to the instructions of the apoptosis detection kit (Best Bio, Shanghai, China), the cells were stained with Annexin V-APC for 10 min and propidium iodide (PI) for 10 min. A BD flow cytometer examined the samples, and FlowJo software (Tree Star Corporation, Ashland, OR, USA) analyzed the data.

Huang Q., Yang Z., Tian J.H., You P.D., Wang J., Ma R., Yu J., Zhang X., Cao J., Cao J, & Wang L.B. (2022). LncSNHG1 Promoted CRC Proliferation through the miR-181b-5p/SMAD2 Axis. Journal of Oncology, 2022, 4181730.

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Cell Cycle and Apoptosis Assay

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The cell cycle assay was performed using propidium iodide fluorescence-activated cell sorting (PI-FACS). After the cell suspension was prepared, the cells were washed with D-Hanks solution, centrifuged, and fixed with 75% alcohol. The fixative was removed, the cells were washed again with D-Hanks, a cell staining solution was added, and cells were then detected using flow cytometry. The cell staining solution consisted of 40× PI (2 mg/mL; Sigma, YSA):100× RNase (10 mg/mL; Fermentas, China):1× D-Hanks = 25:10:1,000.
Apoptosis was detected by Annexin V-APC staining with flow cytometry using an apoptosis detection kit (Bestbio, China) following the manufacturer’s instructions.

Yang R.N., Zhou F.R., Wang H.Y., Wang Q.H., Ji J.L., Huang T., Guo C., Dong Z, & Cao Y.W. (2022). Antitumor activity of RUNX3: Upregulation of E-cadherin and downregulation of the epithelial–mesenchymal transition in clear-cell renal cell carcinoma. Open Life Sciences, 17(1), 1579-1590.

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Apoptosis and Cell Cycle Analysis

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After 48 h of different drugs (0, 25 μM DDP, 25 μM DDP+0.15 μM PGPIPN, 25 μM DDP+1.5 μM PGPIPN and 25 μM DDP+15 μM PGPIPN, respectively) incubations in 6-well culture plates, the cells were digested with trypsin without EDTA and washed twice with pre-cooled PBS. The apoptosis of cells was measured by flow cytometry (FCM) using FITC-conjugated Annexin-V and propidium iodide (PI) from Apoptosis Detection Kit (Bestbio, Shanghai) according to the manufacturer's instructions. The cycle of cell stained with PI was measured by FCM from Cell Cycle Detection Kit (Bestbio, Shanghai) according to the manufacturer's instructions. For cell cycle distribution analysis, per sample analyzed 10,000 cells. The analysis of cell apoptosis and cycle was performed using Flowjo 7.6.1 software. The experiments in cell lines were performed with triplicate, and the experiments were duplicated with primary ovarian cancer cells from six patients.

Guo R., Xu Q., Liu L., Liu H., Liu Y., Wei W, & Qin Y. (2021). Bioactive Hexapeptide Reduced the Resistance of Ovarian Cancer Cells to DDP by Affecting HSF1/HSP70 Signaling Pathway. Journal of Cancer, 12(20), 6081-6093.

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HUVEC Apoptosis Assay by Flow Cytometry

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Cell Counting Kit-8 (Dojindo Molecular Technologies) were used for measurement of the cell viability of HUVECs under various treatments. Apoptosis of HUVECs were analyzed by Apoptosis Detection Kit (BestBio). HUVECs were isolated and resuspended in 100 μL PBS. Annexin V (5 μL) and propidium iodide (10 μL) were add and incubated in dark for about 20 min. Flow cytometer (FACS Calibur, Becton–Dickinson) were used to detect apoptosis of HUVECs.

Cai G.L., Yang Z.X., Guo D.Y., Hu C.B., Yan M.L, & Yan J. (2021). Macrophages enhance lipopolysaccharide induced apoptosis via Ang1 and NF-κB pathways in human umbilical vein endothelial cells. Scientific Reports, 11, 2918.

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Cell Proliferation and Apoptosis Assay

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CCK8 assay was used to detect cell proliferation. The treated HUVECs were added with 10c CCK8 solution (Bestbio, China) per well and incubated at 37 °C for 4 h, followed by measurement of the absorbance at 450 nm with an enzyme label for five consecutive days. An apoptosis detection kit (Bestbio, China) was used to detect apoptosis in treated HUVECs using flow cytometry according to the manufacturer’s instructions.

Zhang X., Leng S., Liu X., Hu X., Liu Y., Li X., Feng Q., Guo W., Li N., Sheng Z., Wang S, & Peng J. (2024). Ion channel Piezo1 activation aggravates the endothelial dysfunction under a high glucose environment. Cardiovascular Diabetology, 23, 150.

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Synthesis and Characterization of Functionalized Nanoparticles

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Silver nitrate (AgNO₃), gold (III) chloride trihydrate (HAuCl₄ · 3H₂O), trisodium citrate dehydrate (Na₃Ct · 2H₂O), N-hydroxy succinimide (NHS), and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) were obtained from Sigma-Aldrich, USA. SH-PEG-COOH was purchased from Shanghai Xibao Medpep Co. The Cell Counting Kit-8 assay (CCK-8) and an apoptosis detection kit were obtained from BestBio Co. Hoechst/PI was purchased from Solarbio Science & Technology Co. 4T1 cells were available from the cell collection of the China Center for Type Culture Collection. Other cell culture reagents were obtained from Gibco. Ltd. Phosphate-buffered solution (PBS) (pH 7.4, 0.1 M) was prepared with 0.1 M Na₂HPO₄, 0.1 M KH₂PO₄, and 0.1 M KCl and was used as the working buffer solution.

Zhang A.W., Guo W.H., Qi Y.F., Wang J.Z., Ma X.X, & Yu D.X. (2016). Synergistic Effects of Gold Nanocages in Hyperthermia and Radiotherapy Treatment. Nanoscale Research Letters, 11, 279.

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Apoptosis Analysis by Flow Cytometry

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Cells for apoptosis analysis were treated according to the procedure as described in cell cycle analysis section. An apoptosis detection kit (BestBio) was employed to analyze apoptotic distribution of the cells. PS (phosphatidylserine) usually found on the inside of the cell membrane and nuclear DNA were used as the biomarkers for the assay. Simultaneous quantification of vital and apoptotic cells was achieved by using dual dying (Annexin V‐FITC (fluorescein isothiocyanate) and PI, targeting PS and nuclear DNA, respectively). Briefly, cells of 10⁶ were collected and washed twice with PBS, followed by being resuspended in 400 µl of binding buffer with the addition of Annexin V‐FITC (5 µl) and PI (10 µl). The resulting mixtures were kept in dark at 4°C for 10 min, and the cells were loaded on the flow cytometer for analysis. The result was analyzed and visualized using CellQuest‐Flow Cytometry Software (Becton Dickinson).

Qin X., Wang X., Xu K., Yang X., Wang Q., Liu C., Wang X., Guo X., Sun J., Li L, & Li S. (2022). Synergistic antitumor effects of polysaccharides and anthocyanins from Lycium ruthenicum Murr. on human colorectal carcinoma LoVo cells and the molecular mechanism. Food Science & Nutrition, 10(9), 2956-2968.

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