The cytotoxicity test was performed after 3, 5 and 7 days using the Cytotoxicity Detection KitPLUS (LDH) (#04744926001; Roche, France) according to the manufacturer’s instructions. This assay is based on the measurement of lactate dehydrogenase (LDH) activity released from the cytosol of damaged cells. Three controls were included: background control (assay medium), low control (untreated cells corresponding to the control condition) and high control (a positive control where a maximum of LDH is released due to cell lysis). The absorbance was read on a spectrophotometer at 490 nm (Varioskan® Flash, Thermo Scientific, Illkirch Graffenstaden, France). To determine the experimental absorbance values, the average absorbance values of triplicate samples and controls were calculated and subtracted from the absorbance values of the background control. The percentage of cytotoxicity was determined relative to the value of the high control (fixed to 100%).
Cytotoxicity detection kitplus ldh
Manufactured by Roche
Sourced in Germany, United States, Switzerland, United Kingdom, France
The Cytotoxicity Detection KitPLUS (LDH) is a colorimetric assay that quantitatively measures the release of lactate dehydrogenase (LDH) from damaged cells. It provides a simple and reliable method for determining cell cytotoxicity and cell-mediated cytolysis.
Automatically generated - may contain errors
Lab products found in correlation
96 well plate, by Thermo Fisher Scientific (8 mentions)
Adi eks 810a, by Enzo Life Sciences (5 mentions)
Diff quik, by Sysmex (4 mentions)
M per, by Thermo Fisher Scientific (4 mentions)
Varioskan flash, by Thermo Fisher Scientific (3 mentions)
Varioskan, by Thermo Fisher Scientific (3 mentions)
96 well plate, by Corning (3 mentions)
Polarstar omega, by BMG Labtech (2 mentions)
Polarstar omega plate reader, by BMG Labtech (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Elisa reader, by Dynex (2 mentions)
Sorafenib, by LC Laboratories (2 mentions)
96 well plate, by Sarstedt (2 mentions)
Fetal bovine serum (fbs), by Equitech-Bio (1 mentions)
Dulbecco s phosphate buffered saline, by Nacalai Tesque (1 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions)
Paraformaldehyde phosphate buffer solution, by Fujifilm (1 mentions)
Penicillin streptomycin, by Nacalai Tesque (1 mentions)
Fxcycle pi rnase staining solution, by Thermo Fisher Scientific (1 mentions)
Cellrox, by Thermo Fisher Scientific (1 mentions)
141 protocols using cytotoxicity detection kitplus ldh
1
Cytotoxicity Assessment of Cell Lines
2
Cytotoxicity and Apoptosis Assays in Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco’s modified eagle’s medium nutrient mixture F-12 HAM (DMEM/F-12) was obtained
from SIGMA (Tokyo, Japan). Fetal bovine serum (FBS) was purchased from Equitech-Bio, Inc.
(Kerrville, TX, U.S.A.). Penicillin/streptomycin, Dulbecco’s phosphate-buffered saline
(PBS) and menadione were purchased from Nacalai tesque (Kyoto, Japan). Camptothecin,
nocodazole, 4% paraformaldehyde phosphate buffer solution (4% PFA), N-acetyl-l -cysteine
(NAC), catalase, mercury (II) chloride (HgCl2), hydrogen peroxide
(H2O2) were from Wako (Osaka, Japan), FxCycle PI/RNase staining
solution and CellROX from Thermo Fisher Scientific (Carlsbad, CA, U.S.A.) and Accumax from
Innovative Cell Technologies (San Diego, CA, U.S.A.). Methylmercury (II) chloride standard
(MeHg) and cadmium chloride 2.5-hydrate (CdCl2) were obtained from Kanto
Chemical (Tokyo, Japan). Cell counting Kit-8 was from Dojindo (Kumamoto, Japan).
Cytotoxicity detection KitPLUS (LDH) and Cell proliferation ELISA, BrdU (colorimetric)
were purchased from Roche (Basel, Switzerland). Amplite fluorimetric Caspase 3/7 assay kit
was from AAT Bioquest (Sunnyvale, CA, U.S.A.).
from SIGMA (Tokyo, Japan). Fetal bovine serum (FBS) was purchased from Equitech-Bio, Inc.
(Kerrville, TX, U.S.A.). Penicillin/streptomycin, Dulbecco’s phosphate-buffered saline
(PBS) and menadione were purchased from Nacalai tesque (Kyoto, Japan). Camptothecin,
nocodazole, 4% paraformaldehyde phosphate buffer solution (4% PFA), N-acetyl-
(NAC), catalase, mercury (II) chloride (HgCl2), hydrogen peroxide
(H2O2) were from Wako (Osaka, Japan), FxCycle PI/RNase staining
solution and CellROX from Thermo Fisher Scientific (Carlsbad, CA, U.S.A.) and Accumax from
Innovative Cell Technologies (San Diego, CA, U.S.A.). Methylmercury (II) chloride standard
(MeHg) and cadmium chloride 2.5-hydrate (CdCl2) were obtained from Kanto
Chemical (Tokyo, Japan). Cell counting Kit-8 was from Dojindo (Kumamoto, Japan).
Cytotoxicity detection KitPLUS (LDH) and Cell proliferation ELISA, BrdU (colorimetric)
were purchased from Roche (Basel, Switzerland). Amplite fluorimetric Caspase 3/7 assay kit
was from AAT Bioquest (Sunnyvale, CA, U.S.A.).
3
Quantifying Cell Membrane Integrity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LDH measurement is a widely utilized method for assessing toxicity and cell membrane integrity.22 (link) LDH measurement was performed in untreated (n=2), SSPCF-treated (n=3) and 1% Triton X-100 in saline solution-treated tissues (positive control, Fluka Biochemika, NJ, USA, n=2) by Cytotoxicity Detection KitPLUS (LDH) (Roche, St. Louis, MO, USA) following manufacturer’s instructions.
4
Screening of Phenolic Compounds for Anti-Amyloidogenic Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lawsone and caffeic acid were purchased from Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan). Juglone and syringic acid were purchased from Sigma (St. Louis, MO, USA). Vanillic acid was purchased from Kanto Chemical Co., Inc. (Tokyo, Japan). Trans-cinnamic acid was purchased from Nacalai Tesque, Inc. (Kyoto, Japan). Chlorogenic acid and ferulic acid were isolated in previous studies from Heynea trijuga [48 ] and Ligusticopsis wallichiana [49 (link)], respectively. Thioflavin T was purchased from Wako Pure Chemical Industry Co. Ltd. (Osaka, Japan), PROTEOSTAT® Protein aggregation assay was purchased from Enzo Life Sciences, Inc. (New York, NY, USA) and the Cytotoxicity Detection Kit Plus (LDH) was purchased from Roche Diagnostic GmbH (Mannheim, Germany).
5
Seletalisib Cytotoxicity in Keratinocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytotoxicity towards different doses of seletalisib was tested by measuring the activity of lactate dehydrogenase (LDH) released from keratinocyte cultures, using Cytotoxicity Detection Kit Plus-LDH (Roche Diagnostics, Milan, Italy) and following the manufacturer’s instructions.
6
Cytotoxicity Evaluation of BLE Extract
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cytotoxicity of BLE extract on the DU145 cell line was determined by measuring the activity of lactate dehydrogenase (LDH) released from the cytosol of damaged cells using a Cytotoxicity Detection Kit PLUS (LDH) (Roche Diagnostics GmbH). The assay was conducted following the manufacturer's instructions. Briefly, medium supernatants of control and treated cells were collected and equal volume of reaction mixture was added. The reaction mixtures were incubated for 20 min at room temperature in the dark. Following incubation, the absorbance was determined at 490 nm using an ELISA plate reader. The percentage of cytotoxicity was calculated as: (exp.value - spontaneous control release) ÷ (max.control release - spontaneous control release) ×100.
7
Cytotoxicity Evaluation of Compounds in Breast and Skin Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human epiterial breast cells MCF10A and human keratinocytes HaCaT (1 × 104/mL) cells were placed on 96-well plates (Nunc, Roskilde, Denmark). The next day, the cells were exposed to increasing concentrations of compounds 3 , 8 and 10 in 0.1, 0.25, 0.5, 1, 2.5, 5, 10 and 25 μM in fresh culture medium. Cytotoxicity was estimated based on the measurements of cytoplasmic lactate dehydrogenase (LDH) activity released from the damaged cells after the 72 h exposure to the compounds. The LDH assay was performed according to the manufacturer’s instructions (Cytotoxicity Detection KitPLUS LDH, Roche, Mannheim, Germany). Absorbance was measured using a microplate at two different wavelengths: “measurement wavelength” (492 nm) and “reference wavelength” (690 nm), using a microplate [61 (link)].
8
Quantifying Cell Cytotoxicity via LDH
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total LDH activity was assessed using the Cytotoxicity Detection Kit PLUS (LDH) (Roche Diagnostics,Florham Park, NJ). Different numbers of cells were plated in 96-well plates and incubated (37°C, 5%CO2, humidified incubator) for 2 hours for their attachment. LDH activity from lysed cells was measured as described [14 (link)].
9
Cytotoxicity Evaluation of FRX on Melanoma and Skin Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Optimized amounts of malignant melanoma cells (as above), normal human keratinocytes HaCaT (1 × 104/mL), and normal human melanocytes HEMa-LP (5 × 103/mL) cells were placed on 96-well plates (Nunc). The next day, the cells were washed in PBS and then exposed to increasing concentrations of FRX in fresh culture medium. The cytotoxicity was estimated based on the measurement of cytoplasmic lactate dehydrogenase (LDH) activity released from damaged cells after 72 h of exposure to FRX. The LDH assay was performed according to the manufacturer’s instruction (Cytotoxicity Detection KitPLUS LDH) (Roche). More information about the steps of the LDH assay has been described earlier [37 (link),38 (link)].
10
Cytotoxicity Assay of MCMV-Specific CD8 T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The sorted pulmonary IV CD45− CD8 T lymphocytes were co‐cultured with 200 pfu/ml heat‐inactivation MCMV for 7 days and used as effector cells. The gB‐transfected autologous SP2/0 cells (H‐2d) were used as target cells. Effector cells and target cells were titrated in U‐bottom 96‐well plates at effector‐target cell ratios of 50:1, 25:1 and 12.5:1; thereafter, 1 × 104 target cells were added and incubated at 37°C for 72 h. Cytotoxicity was determined by measuring the amount of lactate dehydrogenase (LDH) in the supernatant with the Cytotoxicity Detection KitPLUS (LDH) (Roche) according to the manufacturer's instructions. To determine the % cell‐mediated cytotoxicity, calculate the average absorbance of the repeated wells subtract the background, then substitute the resulting values in the following equation:
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!