Cell death elisa kit

Manufactured by Roche

Sourced in Germany, United States

The Cell Death ELISA Kit is a quantitative assay designed to detect and measure cell death in cell culture samples. It utilizes the enzyme-linked immunosorbent assay (ELISA) technique to quantify the amount of histone-complexed DNA fragments in the cytoplasmic fraction of cell lysates, which are indicative of apoptosis or necrosis.

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Spelling variants of this product

Cell Death Detection ELISA kit (275 citations) Cell Death ELISA (23 citations) Cell Death Detection ELISA Plus Assay kit (5 citations) Cell Death Detection enzyme-linked immunosorbent assay (ELISA) kit (5 citations) ELISA‐based cell death detection kit (5 citations) Cell Death Detection ELISA assay kit (4 citations) ELISAs (2 citations) Cell Death Plus ELISA kit (2 citations) ELISA kit for Cell Death Detection (2 citations) Component NO.2 of cell death detection ELISA kit (2 citations)

38 protocols using cell death elisa kit

Cytosolic Nucleosome Quantification

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DNA fragmentation was quantified by measuring the cytosolic nucleosomes using a Cell Death ELISA kit (Roche Diagnostics GmbH, Indianapolis, United States).

Alhusaini A.M., Fadda L.M., Alanazi A.M., Sarawi W.S., Alomar H.A., Ali H.M., Hasan I.H, & Ali R.A. (2022). Nano-Resveratrol: A Promising Candidate for the Treatment of Renal Toxicity Induced by Doxorubicin in Rats Through Modulation of Beclin-1 and mTOR. Frontiers in Pharmacology, 13, 826908.

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Quantifying NETs and MPO/DNA Complexes

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NETs in plasma were determined as NE/DNA complexes in human samples and MPO/DNA complexes in mouse samples. For the human ELISA we used the precoated and blocked plates of the Hycult human NE ELISA (HK319-02). Undiluted plasma samples (50 μl) were incubated for 2h at room temperature with 350 rpm agitation and washed three times with PBS-0.05% Tween (PBS-T). The anti-DNA-POD antibody (Cell Death Detection ELISA Kit, Roche) was diluted 1:100, and the plate was incubated for 2h at room temperature, followed by five washes with PBS-T and incubation with TMB substrate. Signal was acquired at 450 nm.
For the mouse ELISA, the biotinylated primary mouse anti-MPO antibody (1μg/ml final concentration, HM1051BT, Hycult) was coated onto a streptavidin coated plate from the Cell Death Detection ELISA Kit (Roche) at 4°C overnight, followed by three washes with PBS-T. The plates were subsequently blocked for 2h with 1 % BSA in PBS and 50 μl undiluted mouse serum was added to wells. The plate was incubated for 2h at room temperature with agitation (300 rpm on a plate shaker), followed by three washes with PBS-T and addition of 50 μl per well of anti-DNA-POD from Roche cell death ELISA kit (1:100). The plate was incubated for two hours with agitation at room temperature, washed five times with PBST and developed with ABTS.

Knackstedt S.L., Georgiadou A., Apel F., Abu-Abed U., Moxon C.A., Cunnington A.J., Raupach B., Cunningham D., Langhorne J., Krüger R., Barrera V., Harding S.P., Berg A., Patel S., Otterdal K., Mordmüller B., Schwarzer E., Brinkmann V., Zychlinsky A, & Amulic B. (2019). Neutrophil extracellular traps drive inflammatory pathogenesis in malaria. Science immunology, 4(40), eaaw0336.

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Anoikis Assay for Cell Death

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Anoikis (anchorage-independent viability) assay was performed by culturing cells in suspension on polyhema-coated (Sigma-Aldrich) plates, harvested after 24 or 48 h, and cell death assessed using the Cell Death ELISA Kit (Roche) according to the manufacturer’s instructions.

Byerly J.H., Port E.R, & Irie H.Y. (2020). PRKCQ inhibition enhances chemosensitivity of triple-negative breast cancer by regulating Bim. Breast Cancer Research : BCR, 22, 72.

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Quantifying Cytosolic Histone-Bound DNA Fragments

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We measured the levels of cytosolic histone-bound DNA fragmen-ts using a Cell Death ELISA kit (Roche Diagnostics, Mannheim, Germany), as described by the manufacturer.

Woo C.Y., Baek J.Y., Kim A.R., Hong C.H., Yoon J.E., Kim H.S., Yoo H.J., Park T.S., Kc R., Lee K.U, & Koh E.H. (2019). Inhibition of Ceramide Accumulation in Podocytes by Myriocin Prevents Diabetic Nephropathy. Diabetes & Metabolism Journal, 44(4), 581-591.

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Cytoplasmic Histone-Associated DNA Fragments Assay

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A photometric enzyme immunoassay (Cell Death ELISA kit, #11 544 675 001, Roche Applied Science, Penzberg, Germany) was used for quantitative in vitro determination of cytoplasmic histone-associated DNA fragments (mono- and oligonucleosomes). Briefly, HuT-78 and MJ cell lines were seeded at density of 0.3 × 10⁶ cells/mL, cultivated for 24 h until entering the exponential growth phase, and exposed to 80 µM MCRM or 20 µM ERF or the combination thereof. After 72 h incubation period, cells were counted, and the immunoassay was performed according to the manufacturer’s manual. The absorbance of the reaction product was measured at λ = 405 nm (490 reference wavelength) against a substrate solution blank.

Trochopoulos A.G., Ilieva Y., Kroumov A.D., Dimitrova L.L., Pencheva-El Tibi I., Philipov S., Berger M.R., Najdenski H.M., Yoncheva K., Konstantinov S.M, & Zaharieva M.M. (2022). Micellar Curcumin Substantially Increases the Antineoplastic Activity of the Alkylphosphocholine Erufosine against TWIST1 Positive Cutaneous T Cell Lymphoma Cell Lines. Pharmaceutics, 14(12), 2688.

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Oxidative Stress-Induced Cell Death Assay

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Cytoplasmic histone-associated DNA fragments were detected using the cell death ELISA kit from Roche Diagnostics (Indianopolis, IN). Briefly, after incubation with oxidative stressors, cells were lysed and centrifuged at 200 g (10 min), and immunogen and supernatant mix was applied on the ELISA well and incubated for 2 h at room temperature. After washing, substrate solution was added to the wells and incubated until color change that was measured at 405 nm. After background absorbance subtraction, DNA fragment enrichment factor of each sample was calculated as a ratio of sample absorbance to untreated control.

Francelin C., Mitter S.K., Qian Q., Barodia S.K., Ip C., Qi X., Gu H., Quigley J., Goldberg M.S., Grant M.B, & Boulton M.E. (2021). BACE1 Inhibition Increases Susceptibility to Oxidative Stress by Promoting Mitochondrial Damage. Antioxidants, 10(10), 1539.

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Apoptosis Induction Assay by AELE

Cited 1 time

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Cells were seeded and incubated overnight at a density of 1 × 10⁵ cells/ml in 24-well plates. Triplicates of wells treated with two increasing concentrations of AELE for 24 h, were prepared and then compared to untreated control cells. A positive control well, treated with 100 μM of etoposide (Abcam), was also included. Cells were extracted and lysed with incubation buffer, using the Cell Death ELISA kit (Roche), before isolation of fragmented cytosolic DNA. The procedure was then completed as previously described by Ghanem et al. [36 (link)]

Ammoury C., Younes M., El Khoury M., Hodroj M.H., Haykal T., Nasr P., Sily M., Taleb R.I., Sarkis R., Khalife R, & Rizk S. (2019). The pro-apoptotic effect of a Terpene-rich Annona cherimola leaf extract on leukemic cell lines. BMC Complementary and Alternative Medicine, 19, 365.

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Quantification of Cell Viability and Apoptosis in HCC Cells

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For MTT assay and DNA fragmentation detection, HCC cells were seeded the day before drug treatment at a density of 5 × 10³~1 × 10⁴ cells/well in 0.1 mL culture medium onto the 96‐well tissue culture plates. The cells were exposed to various concentrations of indicated drug for 24 or 48 h. Cell viability was measured by the 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide (MTT). DNA fragmentation was assayed by cell death ELISA kit (Roche Life Science, Mannheim, Germany) according to the manufacturer's instructions. For sub‐G1 analysis, 2 × 10⁵ HCC cells were seeded on six‐well plates. After 24 h of drug treatment, cells were harvested by trypsin digestion and fixed in 70% ethanol overnight. The cells were stained with 10 μg·mL⁻¹ propidium iodide for half an hour. The percentage of cells in sub‐G1 phase was quantified by flow cytometry.

Hsieh F., Chen Y., Hung M., Chu P., Tsai M., Chen L., Hsiao Y., Shih C., Chang M., Chao T., Shiau C, & Chen K. (2017). Palbociclib induces activation of AMPK and inhibits hepatocellular carcinoma in a CDK4/6‐independent manner. Molecular Oncology, 11(8), 1035-1049.

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Quantification of Cellular Apoptosis

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Apoptosis was analyzed as previously described (Liu et al., 2016 (link)). Briefly, cells were seeded at a density of 10⁶ cells in a 6-cm dish overnight. The cells were then incubated with doxorubicin or tunicamycin for various time points. Cells were harvested with 250 µl lysis buffer (Cell Death ELISA Kit; Roche). The lysate was centrifuged at 200 g for 5 min, and 20 µl supernatant was incubated with immunoreagent containing 4 µl anti–histone-biotin, 4 µl anti–DNA-POD, and 72 µl incubation buffer (Cell Death ELISA Kit) at room temperature for 2 h in the streptavidin-coated well of the microplate. The reaction mixture was removed, and the well was washed three times with 250 µl of incubation buffer. Next, the well was incubated with 100 µl ABTS substrate solution (Cell Death ELISA Kit) at room temperature for 5 min, followed by absorbance measurement at 405 nm.

Chen F.Y., Huang M.Y., Lin Y.M., Ho C.H., Lin S.Y., Chen H.Y., Hung M.C, & Chen R.H. (2019). BIK ubiquitination by the E3 ligase Cul5-ASB11 determines cell fate during cellular stress. The Journal of Cell Biology, 218(9), 3002-3018.

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Quantification of DNA Fragmentation in PC12 Cells

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The quantification of DNA fragmentation was determined by Cell Death ELISA Kit (Roche Applied Sciences, Basel, Switzerland) according to the manufacturer's protocol. Briefly, PC12 cells were seeded on a 96-well culture plate at a density of 2 × 10⁴ cells/well. At the end of the treatment, the cells were resuspended and incubated in 200 μL of lysis buffer for 30 min at the room temperature. After being centrifuged to remove nuclei and cellular debris, 20 μL of the supernatant of each sample was transferred to a streptavidin-coated microplate and incubated for 2 h at the room temperature with an immunoreagent containing two monoclonal antibodies, anti-histone (biotin-labelled) and anti-DNA (peroxidase-conjugated). After being washed three times with incubation buffer, each well was treated with the chromogen 2,2′-azinobis(3-ethylbinzthiazoline-6-sulfonic acid) as a substrate and then incubated for 15 min. The intensity of the color that developed was measured at 405 nm, while that at 490 nm was used as a blank (reference wavelength). The results were expressed as percentage of nontreated control.

Lee M.K., Lu Y., Di L.Q, & Xu H.Q. (2014). Protection of Tong-Sai-Mai Decoction against Apoptosis Induced by H2O2 in PC12 Cells: Mechanisms via Bcl-2-Mitochondria-ROS-INOS Pathway. Evidence-based Complementary and Alternative Medicine : eCAM, 2014, 371419.

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