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C0017

Manufactured by Beyotime
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Sourced in China

The C0017 is a laboratory equipment designed for general scientific applications. It is a versatile and reliable device that can be used for a variety of tasks in a research or laboratory setting.

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Lab products found in correlation

Microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions) Ldh cytotoxicity assay kit, by Beyotime (1 mentions) S0053, by Beyotime (1 mentions) P0013, by Beyotime (1 mentions) Dmi4000 b system, by Leica (1 mentions)

Close spelling variants of this product

LDH) (c0017; LDH) cytotoxicity assay kit (C0017)

11 protocols using c0017

1

Evaluating CAR-T Cell Cytotoxicity Using LDH Assay

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LDH based assay was conducted to evaluate the cytotoxic effects of CAR-T cells [30 (link)]. Tumor cells (1 x 105 cells /well) were added to the 96-well plates, and then according to the effect of target ratio 1:1, 5:1, and 20:1, c-Met CAR-T, CD19 CAR-T, and non-transduced T cells were added. Next, the incubation was performed at 37°C and CO2 (5%) post 6 hours of culture.And LDH detection reagent was added (C0017, Beyotime, China), the optical density was recorded at 490 nm wavelength.
The mean value of the three wells was substituted into the following formula to calculate the killing efficiency % = (the measured hole – effector cells natural release OD value – target cells natural release OD value + blank control OD value)/(the maximum release hole OD value of the target cells – target cells natural release OD value).
Min J., Long C., Zhang L., Duan J., Fan H., Chu F, & Li Z. (2022). c-Met specific CAR-T cells as a targeted therapy for non-small cell lung cancer cell A549. Bioengineered, 13(4), 9232-9248.
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2

Quantification of Oxidative Stress Markers

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The production of SOD and MDA in lung tissues and BEAS-2B cells were determined with their commercial kits from Beyotime (S0088 and C0017, Shanghai, China). All operations of this assay were following the instruction. The ROS level was detected using its commercial kit from Shanghai Jianglai Biological Technology Co., Ltd. (Shanghai, China).
Yang N, & Shang Y. (2022). Ferrostatin-1 and 3-Methyladenine Ameliorate Ferroptosis in OVA-Induced Asthma Model and in IL-13-Challenged BEAS-2B Cells. Oxidative Medicine and Cellular Longevity, 2022, 9657933.
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3

Cell Viability and Cytotoxicity Assay

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Cell viability was assessed by Cell Counting Kit (CCK‐8; MCE, HY‐K0301, NJ, US) according to the manufacturer's instructions. Briefly, cells were seeded at a concentration of 4000 cells/200 µL well−1 into 96‐well plates, incubated overnight, and changed to fresh medium with various inhibitors. Following treatment, 10 µL CCK‐8 solution was added and cells were incubated for 4 h at 37 °C. The optical density (OD) value was measured at 450 nm by a microplate spectrophotometer (Thermo Fisher Scientific). After trypsinization of cells, placental blue staining was added, and viable cells were enumerated using a hemocytometer. Three independent experiments were performed, each in triplicates.
PRDX5 protein or siPRDX5 was added in the absence or presence of drugs. After 24 h treatment, lactate dehydrogenase (LDH) release assay was performed using a 2‐p‐iodophenyl‐3‐nitrophenyl tetrazolium chloride/ diaphorase‐based kit (C0017, Beyotime, Jiangsu, CN) according to the manufacturer's instructions.
Wang R., Mi Y., Ni J., Wang Y., Ding L., Ran X., Sun Q., Tan S.Y., Koeffler H.P., Feng N, & Chen Y.Q. (2023). Identification of PRDX5 as A Target for The Treatment of Castration‐Resistant Prostate Cancer. Advanced Science, 11(9), 2304939.
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4

LDH Cytotoxicity Assay Protocol

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The LDH cytotoxicity assay was executed according to the manufacturer’s procedure (Cat# C0017, Beyotime Biotechnology, Shanghai, China).
Hu W., Wang Z., Zhang H., Mahaman Y.A., Huang F., Meng D., Zhou Y., Wang S., Jiang N., Xiong J., Westermarck J., Lu Y., Wang J., Wang X., Shentu Y, & Liu R. (2022). Chk1 Inhibition Ameliorates Alzheimer’s Disease Pathogenesis and Cognitive Dysfunction Through CIP2A/PP2A Signaling. Neurotherapeutics, 19(2), 570-591.
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5

Assessing H2O2-Induced Cell Injury

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The assessment of H2O2‐induced cell injury was performed by detection of LDH released into the medium. For this experiment, PC12 cells were seeded in 96‐well plates (1 × 104 cells per well) for 24 h and underwent various treatments. The leakage of LDH was evaluated by measuring its activity in the culture medium using the LDH cytotoxicity assay kit in accordance with the manufacturer’s instructions (C0017; Beyotime Biotechnology). Absorbance was determined at 490 nm using a microplate reader.
Zhang Y., Wang Z., Yang J., He Y., Wan H, & Li C. (2021). Analogs of imine resveratrol alleviate oxidative stress‐induced neurotoxicity in PC12 cells via activation of Nrf2. FEBS Open Bio, 11(8), 2127-2138.
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6

Evaluation of Oxidative Stress Markers

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The primary cardiomyocytes that received different treatments were collected. Cell lysate (P0013, Beyotime, China) was added to lyse cells on ice for 30 minutes according to the manufacturer’s instruction. The lysed cells were centrifuged at 4° C for 12000 r/minutes for 15 minutes, and the supernatant was collected and measured according to the protocols provided by the manufacturer. MDA, SOD, GSH/GSSG, and lactate dehydrogenase (LDH) were measured by the commercial kits (S0131S, S0101S, S0053, C0017, Beyotime, China) by ELISA methods respectively. For rats in a different group, the abdominal aorta blood was drawn and centrifuged. The SOD activity, GSH/GSSG ratio and LDH level in serum were measured.
Wang W., Dong L., Lv H., An Y., Zhang C., Zheng Z., Guo Y., He L., Wang L., Wang J., Shi X., Li N, & Zheng M. (2024). Downregulating miRNA-199a-5p exacerbates fluorouracil-induced cardiotoxicity by activating the ATF6 signaling pathway. Aging (Albany NY), 16(7), 5916-5928.
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7

Blue Light and PTX Cytotoxicity Assay

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Cells were exposed to blue light and 50 µM of PTX for 24 h, after which the concentrations of LDH in the culture supernatants were detected using an in vitro Toxicology Assay kit (C0017; Beyotime Biotechnology, Jiangsu, China). Data were expressed in terms of cell mortality, according to the instruction manual.
Zhuang J., Liu Y., Yuan Q., Liu J., Liu Y., Li H, & Wang D. (2018). Blue light-induced apoptosis of human promyelocytic leukemia cells via the mitochondrial-mediated signaling pathway. Oncology Letters, 15(5), 6291-6296.
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8

Quantifying Cell Death in Vorganoids

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To quantify the degree of cell death at Vorganoids that formed at different stages, the culture supernatants were changed to serum-free DMEM 24 h before the culture medium was aspirated from each well, and the sediment was removed by centrifugation at 1000 rpm for 5 min according to the manufacturer’s protocol. A kit was used to measure the lactate dehydrogenase (LDH) concentration in each group of cultures according to the manufacturer’s instructions (C0017, Beyotime, China).
Liu F., Xiao J., Chen L.H., Pan Y.Y., Tian J.Z., Zhang Z.R, & Bai X.C. (2024). Self-assembly of differentiated dental pulp stem cells facilitates spheroid human dental organoid formation and prevascularization. World Journal of Stem Cells, 16(3), 287-304.
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9

Histopathological Analysis of Ischemic Injury

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Loss of neuron and pathological changes in hippocampus, cortex, and mesencephalon in cerebral I/R gerbils were presented by Nissl and hematoxylin–eosin (H&E) staining. After euthanasia, the brains were removed and placed in 4% paraformaldehyde for 24 h, then embedded in paraffin and sliced into 5‐μm‐thick sections. The Nissl bodies in neurons were stained purple‐red by the Nissl staining solution (Beyotime, C0017) for 10 min. For H&E staining, slices were dewaxed in water, stained for 5 min with hematoxylin, fractionated for a few seconds in 1% hydrochloric acid ethanol, and stained for 3 min with eosin. All slices were dehydrated and transparent. After being sealed with neutral gum and dried, they were photographed and analyzed using a microscope. Neuronal loss and pathological characteristics of slices were observed using an optical microscope (Leica DMI4000 B system).
Wu Y., Hu C., Li Z., Li F., Lv J., Guo M., Liu X., Li C., Huo X., Chen Z., Yang L, & Du X. (2024). Development of a new cerebral ischemia reperfusion model of Mongolian gerbils and standardized evaluation system. Animal Models and Experimental Medicine, 7(1), 48-55.
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10

Oxalate-Induced Cell Injury Evaluation

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Total intracellular lactate dehydrogenase (LDH) was measured to detect the cell injury using an LDH measuring kit (C0017, Beyotime Biotechnology, Shanghai, China). Cells were seeded in 96-well plates at a concentration of 5 × 103 cells/well, and the medium was replaced with DMEM without FBS, followed by treatment with or without 700 μM oxalate [20 (link)] for 12 h. Then the supernatants were collected and measured according to the instructions to determine the LDH release activity, which was presented as the fold increase over the control group levels.
Jiang H., Gao X., Gong J., Yang Q., Lan R., Wang T., Liu J., Yin C., Wang S, & Liu Z. (2018). Downregulated Expression of Solute Carrier Family 26 Member 6 in NRK-52E Cells Attenuates Oxalate-Induced Intracellular Oxidative Stress. Oxidative Medicine and Cellular Longevity, 2018, 1724648.
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