Dead cell apoptosis kit

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

The Dead Cell Apoptosis Kit is a laboratory reagent used to detect and quantify apoptotic cell death. It provides fluorescent-labeled Annexin V and propidium iodide to identify early and late stage apoptotic cells through flow cytometry analysis.

Automatically generated - may contain errors

Lab products found in correlation

206 protocols using dead cell apoptosis kit

Camptothecin-induced Apoptosis in Smad4 Knockdown Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Beas2B cells with Smad4 knockdown were treated with 10μM camptothecin for 4h then harvested with TrypLE (Life Technologies) and assayed with the Dead Cell Apoptosis Kit (Life Technologies) per the manufacturer's instructions using a FC500 flow cytometer (BD Biosciences, San Jose, CA). A549 Smad4−/− cells were treated with camptothecin or etoposide, were harvested with TrypLE (Life Technologies) then washed with cold PBS. Cells were assayed with the Fixable Viability Dye eFluor®506, Annexin V PE Apoptosis Detection Kit, and Anti-Human/mouse pH2AX (Ser139) PerCP-eFluor® 710 (all from eBioscience) per the manufacturer's protocol (http://media.ebioscience.com/data/pdf/best-protocols/annexin-v-staining.pdf) using a Gallios flow cytometer.

Haeger S.M., Thompson J.J., Kalra S., Cleaver T.G., Merrick D., Wang X.J, & Malkoski S.P. (2015). Smad4 loss promotes lung cancer formation but increases sensitivity to DNA topoisomerase inhibitors. Oncogene, 35(5), 577-586.

+ Open protocol

+ Expand

Apoptosis Analysis of Mock and CAR T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mock and CAR T cells were stained with Annexin V/Propidium iodide (PI) dyes according to the dead cell apoptosis kit protocol (V13242, Life Technologies) before and after co-culture with cancer cells for 24, 48, and 72 h. The percentage of positive cells was assessed using flowcytometry.

Yazdanifar M., Zhou R., Grover P., Williams C., Bose M., Moore L.J., Wu S.T., Maher J., Dreau D, & Mukherjee P. (2019). Overcoming Immunological Resistance Enhances the Efficacy of a Novel Anti-tMUC1-CAR T Cell Treatment against Pancreatic Ductal Adenocarcinoma. Cells, 8(9), 1070.

+ Open protocol

+ Expand

Apoptosis and Cell Viability Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

Apoptosis rates were determined by the Dead Cell Apoptosis Kit (LIFE Technologies) according to the manufacturerˈs protocol. Cell vitality was examined by CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, Wisconsin, USA). Analysis of apoptotic cells was carried out by FACSCalibur™ (BD Biosciences, Franklin Lakes, New Jersey, USA) together with the corresponding software BD CellQuest™ Pro (https://www.bd.com/en-uk/products/molecular-diagnostics/cytometric-analysis-products) and cell vitality was detected using LUMIstar Optima (BMG Labtech, Ortenberg, Germany).

Binder S., Zipfel I., Müller C., Wiedemann K., Schimmelpfennig C., Pfeifer G., Reiche K., Hauschildt S., Lehmann J., Köhl U., Horn F, & Friedrich M. (2021). The noncoding RNA LINC00152 conveys contradicting effects in different glioblastoma cells. Scientific Reports, 11, 18499.

+ Open protocol

+ Expand

Apoptosis and ROS Analysis of Hepatocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

For apoptosis analysis, CM (Neu)‐treated, CM (Neu + PMA)‐treated, CM (Neu + PMA + DNase 1)‐treated, untreated or CM (NETs)‐treated HepG2 (1 × 10⁶ cells/well) and HepG2.2.15 cells were seeded on 6‐well plates. After incubation for 48 h, each group of cells was collected and analyzed according to the instructions of the Dead Cell Apoptosis Kit (Life Technologies, Carlsbad, California, USA) with Annexin V/FITC and PI. Similarly, GST‐treated, GST‐S100A9‐treated, GST‐S100A9 + FPS‐ZM1‐treated or GST‐S100A9 + TAK‐242‐treated neutrophils were placed in 6‐well plates. After incubation for 12 h, the cells were collected, suspended in 200 μL PBS and stained with DCFH‐DA (10 μmol/L, S9687, Selleck, Houston, Texas, USA) for 30 min at 37°C, followed by ROS analysis by flow cytometry. All the experiments were performed thrice.

Zhan X., Wu R., Kong X., You Y., He K., Sun X., Huang Y., Chen W, & Duan L. (2022). Elevated neutrophil extracellular traps by HBV‐mediated S100A9‐TLR4/RAGE‐ROS cascade facilitate the growth and metastasis of hepatocellular carcinoma. Cancer Communications, 43(2), 225-245.

+ Open protocol

+ Expand

Evaluating Cell Proliferation and Survival

Check if the same lab product or an alternative is used in the 5 most similar protocols

BrdU incorporation during DNA synthesis was measured using the Cell Proliferation ELISA kit (Roche). Cell viability was measured with the Cell Titer-Blue Cell Viability Assay kit (Promega). Colorimetric signals were measured on a SpectraMax M5 (Molecular Devices). For each assay, the IC₅₀ was calculated with the Prism 5 software package (GraphPad). For assessment of clonogenic survival, cells were drug-treated under low serum conditions in 12 well plates. After 48 h, cells were harvested in trypsin-EDTA (Life Technologies), diluted 1:4000 in standard growth media and seeded in 10 cm plates, in triplicate. After 10 days of growth, plates were stained with 0.2% crystal violet in 50% MeOH and destained in water. Colonies containing more than 50 cells were scored. For the quantitation of apoptosis, cells were detached and stained with a fluorescent antibody directed against Annexin V with the Dead Cell Apoptosis Kit (Life Technologies). The fraction of stained cells was determined with a FACSAria II flow cytometer (BD) in the Sidney Kimmel Comprehensive Cancer Center Flow Cytometry Core.

Larsen A.R., Bai R.Y., Chung J.H., Borodovsky A., Rudin C.M., Riggins G.J, & Bunz F. (2014). Repurposing the antihelmintic mebendazole as a hedgehog inhibitor. Molecular cancer therapeutics, 14(1), 3-13.

+ Open protocol

+ Expand

Apoptosis Induction Assessment of Compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols

PC-3 cells were seeded at 2 × 10⁶ cells/well density in 6-well plates. On the following day cells were treated with 5 µM of either compound 4e or 4j or with 200 µM of cisplatin (Accord Healthcare Ltd., Middlesex, UK). After 48-hour treatment, cells were collected and a Dead Cell Apoptosis Kit containing AnnexinV-FITC and propidium iodide (Life Technologies-Hungary, Budapest, Hungary) was used according to the manufacturer’s recommendation. Fluorescence intensities of at least 10,000 cells/sample were measured by FACSCalibur™ (BD Biosciences, Franklin Lakes, CA, USA) and data were analyzed by FlowJo V10 software (BD Biosciences, Franklin Lakes, NJ, USA). Experiments were repeated three times using triplicates.

Mótyán G., Gopisetty M.K., Kiss-Faludy R.E., Kulmány Á., Zupkó I., Frank É, & Kiricsi M. (2019). Anti-Cancer Activity of Novel Dihydrotestosterone-Derived Ring A-Condensed Pyrazoles on Androgen Non-Responsive Prostate Cancer Cell Lines. International Journal of Molecular Sciences, 20(9), 2170.

+ Open protocol

+ Expand

Neutrophil-Induced Oxidative Stress and CRC Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The production of ROS in neutrophils of different treatment groups were analyzed using the dichlorodihydrofluorescein diacetate (H2DCFDA) probe (S9687, Selleck, Houston, Texas, USA) by flow cytometry. Neutrophils (Neu) were stimulated with Fn, PMA, Fn + TAK-242, Fn + ML130, or Fn + GSK717 for 6 h, then cells were collected and washed with PBS and then stained with 10 µM H2DCFDA for 30 min at 37 °C. Finally, after washing off residual H2DCFDA, cells were analyzed using flow cytometry (CytoFLEX). To investigate apoptosis of CRC cells, which had been treated with (Neu)-CM, (Neu + Fn)-CM, (Neu + Fn + DNase I)-CM, (Neu + PMA)-CM or NETs for 48 h, Dead Cell Apoptosis Kit (Life Technologies) with Annexin V / FITC and PI was used according to the manufacturer’s recommendation. All the experiments were performed three times.

Kong X., Zhang Y., Xiang L., You Y., Duan Y., Zhao Y., Li S., Wu R., Zhang J., Zhou L, & Duan L. (2023). Fusobacterium nucleatum-triggered neutrophil extracellular traps facilitate colorectal carcinoma progression. Journal of Experimental & Clinical Cancer Research : CR, 42, 236.

+ Open protocol

+ Expand

Cell Viability and Apoptosis Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Dulbeccos’s Modified Eagle Medium (DMEM), Trypsin, Dimethylsulfoxide, DAPI, Sulforhodamine B (SRB), Triton X 100 and Phalloidin-FITC were purchased from Sigma Aldrich (Saint Louis, MO, USA), Fetal Bovine Serum (FBS), antibiotic (penicillin 10,000 U/ml; streptomycin 10,000 mg/ml) and Bovine Serum Albumin (BSA) from Gibco (Life Technologies, India) EDTA, Doxorubicin Hydrochloride (Rubidox, Bergamo), Trichloroacetic acid (Neon), Acetic acid (Synth), Methyl alcohol (Neon), TRIS-base (Inlab Confiança, Brazil), Dead Cell Apoptosis Kit (Life Technology™, Carlsbad, California), Accutase® (Thermo Fisher Scientific, Carlsbad, Califórnia, EUA).

de Franca M.N., Isidório R.G., Bonifacio J.H., dos Santos E.W., Santos J.F., Ottoni F.M., de Lucca Junior W., Scher R., Alves R.J, & Corrêa C.B. (2021). Anti-proliferative and pro-apoptotic activity of glycosidic derivatives of lawsone in melanoma cancer cell. BMC Cancer, 21, 662.

+ Open protocol

+ Expand

Camptothecin-induced Apoptosis in Smad4 Knockdown Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Apoptosis Induction by 3BP in MM Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The induction of apoptosis following the action of 3BP was measured using the Dead Cell Apoptosis Kit with Annexin V FITC (AV) and Propidium iodide (PI) (Life Technologies). MM cells at a concentration of 2.5 × 10⁵ cells per milliliter were treated with the appropriate concentration of 3BP (0 μM, 25 μM, 50 μM and 100 μM) and camptothecin (5 μM) as a positive control. Cells were incubated at 37°C under 5% CO₂ for 2 hrs. Using BD FACSCalibur™, the fluorescence was detected. Cells were classified as follows: viable cells (AV-/PI-), early apoptotic cells (AV+/PI-), late apoptotic cells (AV+/PI+) and necrotic cells (AV-/PI+).

Niedźwiecka K., Dyląg M., Augustyniak D., Majkowska-Skrobek G., Cal-Bąkowska M., Ko Y.H., Pedersen P.L., Goffeau A, & Ułaszewski S. (2016). Glutathione may have implications in the design of 3-bromopyruvate treatment protocols for both fungal and algal infections as well as multiple myeloma. Oncotarget, 7(40), 65614-65626.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!