LDH Assay Kit (Cytotoxicity)

• Biochemical assay

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Biochemical assay - LDH Assay Kit (Cytotoxicity) (AB65393)

Diagram showing the principles of the LDH assay method.

• Enzyme activity assay

PubMed

Enzyme activity assay - LDH Assay Kit (Cytotoxicity) (AB65393)

Salama et al. used LDH Assay Kit ab65393 to investigate cytotoxicity when melanoma cell cultures were treated with a potential therapeutic small molecule YO-2.

The LDH assay kit was used to determine cytotoxicity in B16F10 cells treated with/without YO-2 after 24 h (n = 6).

B16F10 cells (1 Ч 105 cells/well) were seeded in triplicate in 6-well plates (Thermo Fisher Scientific, Lafayette, CO, USA) and were left for 16 h overnight to attach before the treatment started. We treated cells with/without YO-2, or DMSO/PBS controls for 24 h.

• FuncS

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Functional Studies - LDH Assay Kit (Cytotoxicity) (AB65393)

Jurkat T cells were cultured in 96-well plate in 100 μl of culture medium. LDH Assay was performed using 10μl of culture medium using the WST probe. Low control (white bar); High control (black bar).

• FuncS

PubMed

Functional Studies - LDH Assay Kit (Cytotoxicity) (AB65393)

Bafilomycin A1 (BafA1) toxicity was assessed in Human hepatic (Huh7.5) cells after 24 hours at different concentrations (12.5nm, 25nm, 50nm and 100nm) were administered to cells using LDH cytotoxicity assay kit (ab65393). Cytotoxicity was measured by subtracting LDH content in remaining viable cells from total LDH in untreated controls.

Image from Bukong TN et al., PLoS Pathog 10(10), Fig S6. Doi: 10.1371/journal.ppat.1004424. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• FuncS

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Functional Studies - LDH Assay Kit (Cytotoxicity) (AB65393)

Comparison of WST-1 and INT based LDH assays. 3T3 cells were cultured in a 96-well plate in 100 μl of culture medium. The LDH assay was performed using 10 μl of culture medium using WST-1 (Brown bar) and INT (Green bar) methods. The WST-1 based LDH assay is more stable and sensitive than the INT based method.

• Enzyme activity assay

PubMed

Enzyme activity assay - LDH Assay Kit (Cytotoxicity) (AB65393)

Chen et al. used LDH Assay Kit ab65393 to investigate cytotoxicity in a placental model of SARS-CoV-2 infection.

LDH levels in the supernatant of 55F STs during differentiation. n = 3 independent experiments

Supernatants collected on day 3 after infection (or indicated otherwise) were analysed for LDH levels.LDH was measured using the Abcam LDH cytotoxicity kit II (ab65393) according to the manufacturer’s instructions in undiluted supernatants using an LDH standard curve.

• Enzyme activity assay

PubMed

Enzyme activity assay - LDH Assay Kit (Cytotoxicity) (AB65393)

Snyder et al. used LDH Assay Kit ab65393 to investigate cytoxicity in male and female neurons maintained in next-generation Neurobasal-Plus media and subjected to a mechanical stretch-injury (to model traumatic brain injury).

Box plots of post-injury LDH levels from 3 independent culture isolations.

• Enzyme activity assay

PubMed

Enzyme activity assay - LDH Assay Kit (Cytotoxicity) (AB65393)

Mudyanselage et al. used LDH Assay Kit ab65393 to compare the neurotoxicity of OPs, chlorpyrifos-oxon (CPO), and azamethiphos (AZO) and the carbamate pesticide, aldicarb, to undifferentiated versus differentiated SH-SY5Y neuroblastoma cells.

Toxicity of CPO, AZO, and aldicarb to undifferentiated and differentiated SHSY-5Y cells measured using a LDH assay. Undifferentiated (A) or differentiated (B) SHSY-5Y cells were treated with CPO, AZO, or aldicarb and extracellular LDH production was quantified using an LDH activity assay. Absorbance readings were corrected by subtracting the values of blanks, then the resultant values were normalised to the LDH production from a vehicle control, providing LDH production as a percentage value relative to the vehicle control. Readings were taken from five independent experiments with three replicates for each treatment. Results were analysed using one-way ANOVA with Dunnett's multiple comparison tests and expressed as means ± standard error of the mean (SEM). For significance, *** p < 0.001, **** p < 0.0001.

The production of active extracellular LDH in response to pesticide exposures was measured using an LDH assay kit (ab65393, Abcam, Cambridge, UK) according to the manufacturer's instructions. After agent treatment (1–200 μM, or vehicle control), 50 μL of spent media was removed and LDH activity was measured spectrophotometrically at 450 nm (Multiskan Spectrum, Thermo Electron Corporation, Finland). Assays were performed in triplicates, with blank values from a negative control subtracted from test values. IC50 values were obtained from the concentration-response curves and expressed as means ± standard error of the mean (SEM). Experiments were performed with an n-number of at least five.

• Enzyme activity assay

PubMed

Enzyme activity assay - LDH Assay Kit (Cytotoxicity) (AB65393)

Lee et al. used LDH Assay Kit ab65393 to investigate the effect of Korean Red Ginseng extract (RGE) on the PD-1/PD-L1 interaction and its antitumor effects using a humanized PD-1/PD-L1-expressing colorectal cancer (CRC) mouse model. The CD8+ T cell-mediated tumor cell killing effect of RGE was evaluated using murine hPD-L1-expressing MC38 cells and tumor-infiltrating hPD-1-expressing CD8+ T cells isolated from hPD-L1 MC38 tumor-bearing hPD-1 mice.

Cytotoxicity in co-cultured hPD-L1 MC38 cells was detected by an LDH cytotoxicity assay. a co-culture cell system was established using tumor-infiltrating CD8+ T cell as effector cells in conjunction with hPD-L1 MC38 cells as target cells to elucidate the CD8+ T cell-mediated antitumor effect of RGE. The cytotoxicity of hPD-L1 MC38 cells co-cultured with tumor-infiltrated CD8+ T cells was increasingly high in a concentration-dependent manner.

A lactate dehydrogenase (LDH) assay kit (#ab65393, Abcam, Cambridge, UK) was used to measure the cytotoxic activity on target cells via effector cells according to the manufacturer’s instructions. The absorbance of the formazan products was determined at 450 nm using a SpectraMax i3 microplate reader.

• Enzyme activity assay

PubMed

Enzyme activity assay - LDH Assay Kit (Cytotoxicity) (AB65393)

Gaetano et al. used LDH Assay Kit ab65393 to investigate the potential use of nanoparticles based on ACyD8 (ACyD8-NPs) loaded with Idebenone (IDE), a synthetic short-chain analogue of coenzyme Q10, as a potential therapeutic for Parkinson's disease.

Analysis of cytotoxicity on the neuronal SH-SY5Y cells pretreated with IDE/ACyD8-NPs and exposed to 30 μM H2O2 for 72 h. Concentrations in IDE/ACyD8-NPs refers to the actual loading of IDE within NPs. Unloaded ACyD8-NPs (i) and (ii) were tested at the same ACyD8 concentration used for testing the drug-loaded NPs 20 μM and 40 μM, respectively. Data are presented as mean ± S.D. One-way ANOVA with Tukey's multiple comparison test : in (a), all data are not statistically different; in (b), * p < 0.05 (H2O2 + IDE/ACyD8-NPs 2 : 1 40 μM versus H2O2).

For the cytotoxicity analysis, the cells were pretreated with NPs (conditions 1–4) for 4 h and then exposed to 30 μM H2O2. Cytotoxicity was evaluated with the LDH-Cytotoxicity Assay Kit II (Abcam, Cambridge, UK, ab65393) after 72 h.

• Schematic Diagram

Supplier Data

Schematic Diagram - LDH Assay Kit (Cytotoxicity) (AB65393)

Representative image of LDH Assay Kit (Cytotoxicity) ab65393

Components shown from left to right :

- LDH Positive Control

- Substrate Mix

- Stop Solution IV

- Lysis Buffer II

- LDH Assay Buffer

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit.