Ldh cytotoxicity kit

Manufactured by Thermo Fisher Scientific

Sourced in China, United States

The LDH cytotoxicity kit is a laboratory assay used to measure the release of lactate dehydrogenase (LDH) from damaged cells. It provides a quantitative assessment of cell cytotoxicity or cell death.

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Lab products found in correlation

Mil 2, by Miltenyi Biotec (2 mentions) Cytometric bead array, by BD (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Aspirin, by Merck Group (1 mentions) Celecoxib, by Merck Group (1 mentions) Hrp conjugated goat anti rabbit igg, by Cell Signaling Technology (1 mentions) High glucose dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Dispase, by BD (1 mentions) Waymouth s medium, by Thermo Fisher Scientific (1 mentions) Anti actin antibody sc 1616, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated donkey anti goat igg, by Santa Cruz Biotechnology (1 mentions) C terminal gfp tagged open reading frame clone of full length human vegfr2, by OriGene (1 mentions) Ham s f10, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Percoll, by Merck Group (1 mentions) Anti caspase 3, by Cell Signaling Technology (1 mentions) Trypsin, by Merck Group (1 mentions) Dnase, by Merck Group (1 mentions) Glass bottom culture 35mm plates, by Ibidi (1 mentions) Poly l lysine (pll), by Ibidi (1 mentions)

10 protocols using ldh cytotoxicity kit

Cytotoxicity Assay for Activated T Cells

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L929 target cells—IL-2 primed splenocytes (effector cells) from WT and PAG KO mice were co-cultured overnight with L929 cells (target cells). The release of LDH from the lysed target cells was measured by the LDH cytotoxicity kit (ThermoFisher), and cytotoxicity was calculated relative to target cells cultured in the absence of primed T cells. Raji target cells—Murine splenocytes were cultured with 1 µg/ml SEE and 1000 U/ml mIL-2 (Miltenyi) for 72 h, followed by isolation of CD8⁺ T cells using isolation kit (Miltenyi). SEE loaded Raji B cells (target cells) were mixed with CD8⁺ T cells at different ratios, as indicated, and cytotoxicity was tested using LDH cytotoxicity kit (ThermoFisher).

Strazza M., Azoulay-Alfaguter I., Peled M., Adam K, & Mor A. (2021). Transmembrane adaptor protein PAG is a mediator of PD-1 inhibitory signaling in human T cells. Communications Biology, 4, 672.

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Quantifying Inflammatory Mediators in Blister Exudates

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Cell-free cantharidin blister exudates and monocyte culture supernatants were assayed for total protein content using a Bradford assay (Bio-Rad) as per the manufacturer’s instructions.
TGF-β levels were determined using a single-plex sandwich ELISA (Affymetrix eBioscience). The assay was performed according to the manufacturer’s instructions.
Other cytokines were measured using an RnD Luminex Screening Assay (Bio-Techne) or cytometric bead array (BD Biosciences). Samples were run in duplicate and the assays were performed according to the manufacturer’s instructions.
Lactate dehydrogenase (LDH) activity was measured in cell-free cantharidin blister exudates using an LDH cytotoxicity kit (Thermo Scientific) according to manufacturer’s instructions.

De Maeyer R.P., van de Merwe R.C., Louie R., Bracken O., Devine O.P., Goldstein D.R., Uddin M., Akbar A.N, & Gilroy D.W. (2020). Blocking elevated p38 MAPK restores efferocytosis and inflammatory resolution in the elderly. Nature immunology, 21(6), 615-625.

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Tau ELISA on Primary Neurons

Cited 1 time

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Tau enzyme-linked immunosorbent assays were performed on Hank’s balanced salt solution with Ca²⁺ and Mg²⁺ medium that had been incubated for 4 h with 22–23 DIV primary neurons as we described previously (Croft et al., 2017 (link)). The amount of lactate dehydrogenase in the media of cultured neurons was determined as a measure of neuron heath, using an LDH Cytotoxicity Kit from Thermo Fischer Scientific according to the manufacturer’s instructions.

Glennon E.B., Lau D.H., Gabriele R.M., Taylor M.F., Troakes C., Opie-Martin S., Elliott C., Killick R., Hanger D.P., Perez-Nievas B.G, & Noble W. (2020). Bridging integrator 1 protein loss in Alzheimer’s disease promotes synaptic tau accumulation and disrupts tau release. Brain Communications, 2(1), fcaa011.

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Measuring Brain LDH Cytotoxicity

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Lactate dehydrogenase (LDH) release from brain tissue as well as neurons was measured using a LDH cytotoxicity kit (Thermo Scientific, Shanghai, China) according to the manufacturer's instructions. The absorbance was measured at 450 nm wavelength with a microplate reader.

Song N., Song R, & Ma P. (2022). MiR‐340‐5p alleviates neuroinflammation and neuronal injury via suppressing STING in subarachnoid hemorrhage. Brain and Behavior, 12(9), e2687.

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Endothelial Cell Functional Assays

Cited 1 time

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Aspirin, celecoxib, trypsin, DNAse and Percoll were purchased from Sigma-Aldrich (St. Louis, MO). Dispase was purchased from B&D Bioscience (San Jose, CA). 5-(4-Chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethylpyrazole (sc-560) was purchased from EMD Millipore (Billerica, MA). The ELISA Kit for human sVEGFR1 and anti-Flt1 antibody (AF321) were obtained from R&D Systems (Minneapolis, MN). Cleaved poly ADP-ribose polymerase (PARP) antibody, anti-caspase-3 and HRP-conjugated goat anti-rabbit IgG were purchased from Cell Signaling Technology (Danvers, MA). Anti-actin antibody (sc-1616) and HRP-conjugated donkey anti-goat IgG were from Santa Cruz Biotechnology (Santa Cruz, CA). The LDH cytotoxicity kit was purchased from Thermo-Fisher Scientific (Rockford, IL). Ham's F-10, Waymouth’s medium, high Glucose Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were from Life Technologies (Grand Island, NY). C-terminal GFP-tagged open reading frame (ORF) clone of full length human VEGFR2 was purchased from OriGene (Rockville, MD). Recombinant adenovirus expressing FLT, sFLT1-i13, sFLT1-e15a and control virus were generated by the vector Core Facility at the University of Iowa and has been previously described (4 (link), 6 (link)).

Li C., Raikwar N.S., Santillan M.K., Santillan D.A, & Thomas C.P. (2015). Aspirin inhibits expression of sFLT1 from human cytotrophoblasts induced by hypoxia, via cyclo-oxygenase 1. Placenta, 36(4), 446-453.

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Cytotoxicity Assay of CD8 T Cells

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Murine splenocytes were cultured with 1 ug/mL SEE and 1000 U/mL mIL-2 (Miltenyi) for 72 hours, followed by isolation of CD8 T cells using isolation kit (Miltenyi). SEE loaded Raji B cells (target cells) were mixed with CD8 T cells at different ratios, as indicated, and cytotoxicity was tested using LDH cytotoxicity kit (ThermoFisher). In some experiments, CD8 T cells were stained with Lysotracker (ThermoFisher) before mixing with Raji cells, plated on poly-L-lysine (5 μg/ml) coated glass bottom culture 35mm plates (ibidi), and subjected to brief centrifugation prior to imaging. Images were taken with Zeiss 700 confocal microscope and analyzed with ImageJ (NIH).

Peled M., Dragovich M.A., Adam K., Strazza M., Tocheva A.S., Vega I.E, & Mor A. (2018). EF Hand domain family member D2 is required for T cell cytotoxicity. Journal of immunology (Baltimore, Md. : 1950), 201(9), 2824-2831.

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Quantification of Inflammasome Activation

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Primary antibodies were mouse anti-capase-1 (AG-20B-0042; Adipogen), mouse anti-FLAG (F1804; Sigma–Aldrich), mouse anti-actin (66009–1-Ig; ProteinTech), anti-ASC (67824; Cell Signaling Technology), anti-GFP (BM3883; Boster Biotechnology), and anti-GSDMD (209845; Abcam). Mouse IL-18 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Invivogen (BMS618). An LDH Cytotoxicity kit was obtained from ThermoFisher (C20300).

Shen C., Li R., Negro R., Cheng J., Vora S., Fu T.M., Wang A., He K., Andreeva L., Gao P., Tian Z., Flavell R., Zhu S, & Wu H. (2021). Phase Separation Drives RNA Virus-Induced Activation of the NLRP6 Inflammasome. Cell, 184(23), 5759-5774.e20.

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Tau ELISA Assay on Primary Neurons

Cited 1 time

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Tau ELISAs were performed on Hank’s balanced salt solution (HBSS)
without with Ca²⁺ and Mg²⁺ medium that had been incubated
for 4 hours with 22-23 DIV primary neurons as we described previously (Croft et al., 2017 (link)).
Lactate dehydrogenase amounts in the media of cultured neurons was determined as
a measure of neuron heath, using an LDH cytotoxicity kit from Thermo Fischer
Scientific according to the manufacturer’s instructions.

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Cytotoxicity Evaluation of CD8+ T Cells

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Upon achieving a 90% growth of CD8 + T cells, the suspension cells and the transfected DLBCL cells were cocultured in a 3:1 ratio. Upon completion of 48 hours of coculture, the cells were collected for subsequent analysis. Subsequently, T cell cytotoxicity was determined using the LDH cytotoxicity kit (Thermo Fisher, MA, USA) in accordance with the manufacturer's guidelines.

Qin X., Li H., Wu J., Tang W., Li W, & Li K. (2024). PGM5-AS1 Promotes Progression of Diffuse Large B-Cell Lymphoma and Immune Escape by Regulating miR-503-5p. Journal of inflammation research, 17.

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Evaluating Macrophage Responses to Mycobacterial Infection

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Murine macrophages RAW 264.7 infected with Ms_Vec and Ms_Rv1523 for 2 h at 37°C, 5% CO₂ and macrophages cells without infection were used as control. For lactate dehydrogenase (LDH) activity analysis, the culture supernatants from infected cells and control cells without infection were harvested after 24 and 48 h after infection. Following the standard protocols, the LDH activities were assessed using commercially available LDH cytotoxicity kit (Thermo). By using a Griess reagent (Thermo) which measures the stable end product, nitrite, the NO levels in culture supernatant of infected macrophages were determined. An equal volume of 100 µl of Griess reagent and culture supernatant was added to a 96-well plate in duplicate and incubated at room temperature for 15 min. Using standard curve for nitrite, levels of nitrite was estimated after taking absorbance at 540 nm. The results were showed as the mean µmoles of nitrite/sample ± SEM.

Ali S., Ehtram A., Arora N., Manjunath P., Roy D., Ehtesham N.Z, & Hasnain S.E. (2021). The M. tuberculosis Rv1523 Methyltransferase Promotes Drug Resistance Through Methylation-Mediated Cell Wall Remodeling and Modulates Macrophages Immune Responses. Frontiers in Cellular and Infection Microbiology, 11, 622487.

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