Cytotoxicity of PCL-PPB-PCo-7 was further performed using human primary umbilical vein endothelia cells (HUVEC) cultured with vascular cell basal medium containing endothelial cells growth factors (ATCC, Manassas, VA). PCL-PPB-PCo-7 copolymers were added to the cell culture medium when cells were at 70–80% confluency. Aliquots of cell culture medium were taken after 1, 8 and 24 h. LDH levels in the media were measured by LDH-Glo Cytotoxicity Assay kit (Promega, Madison, WI) according to the provided instruction. Medium without copolymers was used a negative control to determine culture medium background. For maximum LDH release control, Triton X-100 with the final concentration of 0.2% was added to the cell culture medium. The % cytotoxicity was calculated as (experimental LDH level – background) divided by (maximum LDH – background).
Ldh glo cytotoxicity assay kit
Manufactured by Promega
Sourced in United States
The LDH-Glo™ Cytotoxicity Assay kit is a laboratory product that measures the activity of lactate dehydrogenase (LDH), an enzyme released from damaged or dead cells. The assay provides a quantitative evaluation of cytotoxicity or cell death in cell-based experiments.
Automatically generated - may contain errors
Lab products found in correlation
Enspire luminometer, by PerkinElmer (3 mentions)
Celltiter glo luminescent cell viability assay kit, by Promega (2 mentions)
Spectramax id3, by Molecular Devices (2 mentions)
White half area assay plates, by Greiner (1 mentions)
Spark 10m plate reader, by Tecan (1 mentions)
Tris hcl, by Merck Group (1 mentions)
Spark cyto plate reader, by Tecan (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Glycerol, by Merck Group (1 mentions)
Glomax explorer multimode microplate reader, by Promega (1 mentions)
Celltiter glo 2, by Promega (1 mentions)
Microplate reader, by Tecan (1 mentions)
Digitonin, by Merck Group (1 mentions)
Mccoy 5a medium, by Merck Group (1 mentions)
Immunocult, by STEMCELL (1 mentions)
Glomax 96 microplate luminometer w dual injectors, by Promega (1 mentions)
Quant it picogreen dsdna reagent, by Thermo Fisher Scientific (1 mentions)
96 well plate, by PerkinElmer (1 mentions)
Temozolomide, by Selleck Chemicals (1 mentions)
Mifepristone, by Merck Group (1 mentions)
25 protocols using ldh glo cytotoxicity assay kit
1
Cytotoxicity Assessment of PCL-PPB-PCo-7
2
Quantification of Cell Cytotoxicity by LDH Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Activity of lactate dehydrogenase (LDH) released from damaged cells into the supernatant was measured using the LDH-Glo Cytotoxicity Assay kit (Promega Corporation), according to manufacturer’s protocol. Cell culture supernatants were diluted 1:5 in LDH storage buffer and the diluted samples (25 µl) were mixed with 25 µl detection reagent in white half area assay plates (Greiner Bio-One). After a 1 h incubation on an orbital shaker at room temperature (RT), protected from light, luminescence was measured with a Tecan Spark 10 M plate reader. Measured relative luminescence unit values (RLUs) are directly proportional to the LDH concentration. Background-subtracted RLU values are plotted as mean ± SD, including individual values. Calculations were done in Microsoft Excel and graphs prepared with GraphPad Prism (Version 8) software.
3
Cell Viability and Cytotoxicity Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell killing activity of DM4 or the ADC SAR408701 analog was measured by CellTiter-Glo Luminescent cell viability assay kit (Promega, G7571). Cells (2.5×103 cells/well in white 96-well plate) were cultured for 12 h prior to treatment with the indicated doses of DM4 or ADC for 96 h at 37°C. Normalized % ATP values were calculated by normalizing luminescence values for buffer (DPBS or DMSO)-treated cells. The LDH-Glo cytotoxicity assay kit (Promega, J2381) was used to measure cell viability in treatment of anti-CEACAM5 CAR-T cells. Control T or CAR-T cells as effector cells were incubated with target cells (5×103 cells/well in 96-well plate) at the indicated E:T ratio for 24 h at 37°C. Controls conducted for the calculation of percent cytotoxicity were included according to the manufacturer’s instructions.
4
NK Cell-Mediated Neuroblastoma Cytotoxicity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs and NK cells collected from healthy donors were purchased from the University of Pennsylvania Human Immunology Core facility. Target neuroblastoma cells (NB-EbC1 or SMS-SAN) were plated for 1 day, at which point PBMCs or NK cells were added immediately after the addition of either an isotype control human IgG1 (BioXCell, #BE0297), D3-GPC2-IgG1, or GD2 targeting dinutuximab antibody. Target cell lysis was measured either in real-time on an RT-CES system or by LDH quantification after 24 hours using an LDH-Glo™ Cytotoxicity Assay Kit (Promega) according to the manufacturer’s instructions. LDH concentrations were calculated from a standard curve and the % cytotoxicity normalized to the concentration of LDH released in the experimental sample compared to that of the GD2 targeting dinutuximab positive control (considered 100% cytotoxicity) according to the below equation:
5
Antibody-Dependent Cell-Mediated Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ADCC assay used the constructed NK92/CD16a stable cell line as effector cells and Rituxan (rituximab, targeting CD20) as a positive control antibody. The antibody concentration was started at 20 µg/mL with tenfold dilution and 10 concentration points. The killing effect on target cells was detected using a lactate dehydrogenase (LDH-Glo™ Cytotoxicity Assay) kit (Promega). The complement of CDC was obtained from normal human serum (NHS) with a final serum concentration of 20%. The starting concentration was 20 μg/mL at tenfold dilution and 8 concentration points. Target cell viability was detected using a CellTiterGlo® Luminescent Cell Viability Assay kit (Promega).
6
Measuring Enterotoxin-Induced LDH Release
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LDH activity was determined in the apical tubule media after enterotoxin exposure using the LDH-Glo Cytotoxicity Assay Kit (Promega, J2381). Medium from the right upper and bottom channels was collected and diluted 1:100 in LDH storage buffer (200mM Tris–HCl (Sigma-Aldrich, T2194), 10% glycerol (Sigma, G5516), 1% BSA (Sigma-Aldrich, A2153) and then processed following the manufacturer’s instructions. Those cells incubated with no enterotoxin-containing media were used as negative controls. White half-volume 96-well plates (Greiner, 675,075) were used to perform the assay. Luminescence was measured using the Spark Cyto plate reader (Tecan Life Sciences) using an integration time of 80 ms.
7
LDH-Glo™ Cytotoxicity Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Drug cytotoxicity was measured by the lactate dehydrogenase (LDH)-Glo™ Cytotoxicity Assay Kit (#J2380, Promega) following manufacturer’s instructions. Luminescence was measured at 400 nm using the GloMax® Explorer Multimode Microplate Reader (#GM3500, Promega).
8
Measuring Cell Viability and ROS Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability was measured as described previously [21 (link), 23 (link)]. Briefly, dissociated GT1-7 cells were distributed into 96-well culture plates at a density of 3 × 104 cells per well in 200 μL of culture medium. After 24 h incubation, cells were treated with or without N-acetylcysteine (NAC: 0–250 μM), carnosine (0–4 mM), or TUDCA (0–100 mM) prior to the addition of NiCl2 and ZnCl2 to the medium. After 24 h exposure, cell viability was quantified using CellTiter-Glo® 2.0 (Promega Corporation, Madison, WI). Cytotoxicity was quantified using an LDH-Glo™ Cytotoxicity Assay kit (Promega Corporation, Madison, WI) after exposure for 24 h.
GT1-7 cells were precultured in black 96-well microplates (3 × 104 cells/well). After incubation for 24 h, cells were incubated with 2 ′,7 ′-dichlorodihydrofluorescein diacetate (DCFHDA, 100 μM), an indicator of ROS, in the absence (control) or presence of NiCl2 and/or ZnCl2 for 2 h. ROS levels were then measured using a microplate reader (Tecan, Kawasaki, Japan) (Ex: 480 nm; Em: 530 nm).
GT1-7 cells were precultured in black 96-well microplates (3 × 104 cells/well). After incubation for 24 h, cells were incubated with 2 ′,7 ′-dichlorodihydrofluorescein diacetate (DCFHDA, 100 μM), an indicator of ROS, in the absence (control) or presence of NiCl2 and/or ZnCl2 for 2 h. ROS levels were then measured using a microplate reader (Tecan, Kawasaki, Japan) (Ex: 480 nm; Em: 530 nm).
9
CAR-T Cell Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CAR-T cells were cultured in Immunocult (STEMCELL Technologies; supplemented with 1% PenStrep) under the following conditions for 24 hours: without treatment, with 250 μM adenosine, or with 10 mM MNA. After pretreatment, CAR-T cells were washed with PBS and cocultured with 20,000 SK-OV-3 cells [ATCC; expanded in McCoy 5A medium (Sigma-Aldrich) supplemented with 10% FBS and 1% PenStrep] in supplemented Immunocult media in triplicate at an effector-to-target ratio of 10:1. SK-OV-3 cells and SK-OV-3 cells lysed with digitonin (0.5 mg/ml; Sigma-Aldrich) were used as negative and positive controls, respectively. Following 24 hours of coculture, supernatants were collected, and lactate dehydrogenase (LDH) was measured according to the manufacturer’s instructions (LDH Glo Cytotoxicity Assay Kit, Promega). LDH supernatant was diluted 1:50 in LDH buffer. Percent killing was measured using the following formula: % killing = corrected killing/maximum killing × 100%, where corrected killing = coculture − T cells alone, and maximum killing = positive control − negative control.
10
Evaluating Cytotoxicity in Enteroid-Derived Monolayers
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!