Cytocompatibility was determined using a commercially available lactate dehydrogenase (LDH) assay kit (Cayman Chemical). Immediately following centrifugation and 10 mins rest, PRP was placed on the substrates. After 2 hrs of incubation in PRP, the surfaces were shaken on a horizontal shaker plate (100 rpm) for 5mins at room temperature. 100 µL of surface-exposed PRP and manufacturer supplied standards were transferred to a 96-well plate. A reaction solution consisting of 96% v/v assay buffer, 1% v/v nicotinamide adenine dinucleotide NAD+1%, v/v lactic acid, 1% v/v INT, and 1% v/v LDH diaphorase was added in equal amounts (1:1) to all the standards and surface-exposed PRP. This was followed by gentle shaking on a horizontal shaker plate (100 rpm) for 30 mins at room temperature. Total time from blood collection to end of study was approximately 3 hrs. The absorbance of this was immediately measured at a wavelength of 490 nm using a plate reader (BMG Labtech).
Ldh assay kit
Manufactured by Cayman Chemical
Sourced in United States
The LDH assay kit is a laboratory product used to quantify the activity of the enzyme lactate dehydrogenase (LDH) in biological samples. LDH is a ubiquitous enzyme found in various tissues and is often used as a marker for cellular damage or cytotoxicity.
Automatically generated - may contain errors
Lab products found in correlation
Protein assay kit, by Bio-Rad (3 mentions)
Microplate reader, by Tecan (2 mentions)
Epoch microplate spectrophotometer, by Agilent Technologies (1 mentions)
Phenol red free media, by Thermo Fisher Scientific (1 mentions)
Versamax, by Molecular Devices (1 mentions)
Imark microplate reader, by Bio-Rad (1 mentions)
Elx800, by Agilent Technologies (1 mentions)
Infinite m200 pro, by Tecan (1 mentions)
Rpmi medium, by Thermo Fisher Scientific (1 mentions)
Fungizone, by Thermo Fisher Scientific (1 mentions)
18 protocols using ldh assay kit
1
Cytocompatibility of Platelet-Rich Plasma
2
Quantifying Cell Injury via LDH Release
3
Quantifying Tissue Necrosis via LDH Assay
4
Platelet Cytotoxicity by LDH Leakage
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Human platelet cytotoxicity was determined by the leakage of lactate dehydrogenase (LDH) from cytosol. Human washed platelets (108/mL) were incubated for 5 min at 37°C with various concentrations of G-Ro and then centrifuged at room temperature for 2 min at 12,000 g. The supernatant was measured by LDH assay kit (Cayman Chemical) at an optical density of 490 nm. LDH leakage is expressed as the percentage of the total enzyme activity in platelets completely lysed with 0.1% Triton X-100.
5
Dose-Dependent Cytotoxicity of Quantum Dots on Brain Slices
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QDs were added to brain slices in a dose-dependent manner (0.01, 0.1 and 1 μM) for 24 h, and viability of each whole brain slice was evaluated. Media beneath pre-cut, 300 μm-thick OWH slices on membrane inserts was changed to fresh, pre-warmed slice culture medium. The QD-MPA, QD-PEG-OH and QD-PEG-OMe were diluted to 1, 0.1, and 0.01 μM by 1× PBS, and 100 μL of suspended particles were carefully dropped onto each slice. Slices without QD treatment served as the non-treated control group (NT), and slices receiving 1% Triton X-100 were considered as the 100%-cell-death, maximum-release group (Tx). The slices were then incubated at 37 °C with constant humidity, 95% air, and 5% CO2 for 24 h. At the end of the exposure time, fresh medium was replaced, and supernatant aliquots were taken at 0, 1, 2, 4, 8, and 24 h timepoints. NT and Tx groups were run for each QD-treated study. Supernatants at each time point were utilized to determine cell death with an LDH assay kit, according to manufacturer instructions (Cayman, USA). The percentage of LDH released in each whole-hemisphere brain slice was quantified by measuring the absorbance intensity of the formazan reactive product, subtracting the intensity of NT groups, and normalizing by the intensity of Tx groups. (n = 3)![]()
6
Histone-Induced Cytotoxicity Assay in Rat Cardiomyocytes
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LDH assay was used to detect the cytotoxicity levels of histones on rat CMs according to the protocol which we described before [33 (link)]. Briefly, supernatants fluids were obtained from CMs exposed to histone, using phenol red-free media (Gibco, Grand Island, NY). The percentage of cytotoxicity (LDH release) from the sample treated with histone was measured compared to LDH content in total lysis fluids induced by 0.1% triton detergent (Sigma-Aldrich, St. Louis, MO). LDH release was measured using LDH assay kit (Cayman Chemical, Ann Arbor, MI) according to the manufacturer's instructions.
7
DIE Modulation of LPS-Induced Inflammation
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RAW 264.7 cells were cultured at a density of 3 × 105 cells/mL in 24-well plates overnight. DIE was added at 12.5 μg/mL to 100 μg/mL for 30 min followed by the administration of 0.1 μg/mL LPS and incubated for 18 h before the supernatant was collected for NO assay using the Griess Reagent. 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was added to each well and left to incubate for 3 h. Formazan crystals of MTT were dissolved with DMSO and quantified using a plate reader (VersaMax, Molecular Devices, San Jose, CA, USA). The wavelengths used for measuring NO and MTT were 540 nm and 560 nm, respectively. Lactate dehydrogenase (LDH) activity was detected in RAW 264.7 cells with an LDH assay kit (Cayman Chemicals, Ann Arbor, MI, United States). Briefly, DIE was treated with RAW 264.7 cells as stated above, with or without LPS. LDH activity was detected according to the manufacturer's instructions.
8
Cytotoxicity and Cytokine Secretion of ANPs
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The cytocompatibility of ANPs and the secretion
of tumor necrosis factor-α (TNF-α) from macrophage-like
cells exposed to ANPs were measured using a lactate dehydrogenase
(LDH) assay kit (Cayman Chemical) and ELISA assay (Thermo Fisher Scientific).
Then, 2.5 × 104 RAW264.7 cells were seeded onto a
48-well plate and cultured in DMEM for 24 h. The culture medium was
then replaced by a medium containing a mixture of BSA and OVA (9:1
(w/w), 50 μg/mL) and ANPs with AR1, AR3, and AR10. After 24
h of incubation at 37 °C, the supernatant was collected from
each well and LDH and ELISA assays were performed according to the
manufacturer’s procedures.
of tumor necrosis factor-α (TNF-α) from macrophage-like
cells exposed to ANPs were measured using a lactate dehydrogenase
(LDH) assay kit (Cayman Chemical) and ELISA assay (Thermo Fisher Scientific).
Then, 2.5 × 104 RAW264.7 cells were seeded onto a
48-well plate and cultured in DMEM for 24 h. The culture medium was
then replaced by a medium containing a mixture of BSA and OVA (9:1
(w/w), 50 μg/mL) and ANPs with AR1, AR3, and AR10. After 24
h of incubation at 37 °C, the supernatant was collected from
each well and LDH and ELISA assays were performed according to the
manufacturer’s procedures.
9
Quantification of LDH Activities
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Lactate dehydrogenase (LDH) activities in media were measured using an LDH assay kit (Cayman, Ann Arbor, MI, USA). Briefly, 20S-PPD-treated media were incubated for 30 min at room temperature with an assay buffer containing LDH diaphorase, lactic acid, NAD+, and tetrazolium salt. Absorbance at 490 nm was then measured using a microplate reader (Tecan).
10
Quantifying C2C12 Cell Death
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C2C12 cells were plated on polyD-lysine coated plates/wells and cultured in differentiation media (method described above). After the 5th (AR100Q-SIRT3 overexpressing or AR100Q-EV lines only) or 10th day of treatment the LDH levels were quantified with an LDH assay kit (Cayman Chemical, 601170) using the manufacturer’s protocol. Samples of media were taken from the same well at the end of the treatment period (for experimental sample reading) and after inducing cell death by adding 10% Triton X-100 to media (for 100% cell death control reading). At the end of the assay, samples were evaluated in a colorimetric plate reader (iMarkTM microplate reader, Bio-Rad) at 450 nm. Data generated for LDH concentrations for the experimental samples were compared to the LDH values from the “100% cell death control” to determine the percent cell death.
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