Target/Analyte:
ROS

Short Name ROS
Description Reactive Oxygen Species

Method/Kits with this Target/Analyte
Method/Kit Description
Human Bone Panel I Kit (MSD: K15146C) The Human Bone Marker Panel I Kit provides assay-specific components for the quantitative determination of both natural and recombinant human osteoprotegerin (OPGN), sclerostin (SOST), and alkaline phosphatase (ALP) in serum and plasma
Mass spectrometry Mass spectrometry
LC-MS/MS Determination of analyte concentration by LC-MS/MS
ProcartaPlex assay Cytokine detection assay
Single-Use RED Plate with Inserts (Thermo Scientific: 2034.6) and LC-MS Single-Use RED Plate with Inserts (Thermo Scientific: 2034.6) followed by mass spectrometry for quantification of compound and determination of: % Free = (Concentration buffer chamber/Concentration plasma chamber) × 100% and % Bound = 100% - % Free.
Corrosivity Assessment | PrestoBlue Cell Viability (ThermoFisher: A13261, A13262) An assessment of compound corrosion based on processing data from the PrestoBlue Cell Viability assay.
BODIPY 581/591 C11-Lipid Peroxidation Sensor Measure lipid peroxidation with BODIPY C11 sensor
DHE Reactive oxygen species chemical probe
HCS LipidTOX Red Neutral Lipid Stain (Thermo: H34476) The intracellular accumulation of neutral lipids, steatosis, is often triggered by drugs that affect the metabolism of fatty acids and/or neutral lipids. HCS LipidTOX™ Red neutral lipid stain was developed to characterize the potentially toxic effects of compounds on lipid metabolism in mammalian cell lines. The LipidTOX™ neutral lipid stain has an extremely high affinity for neutral lipid droplets and can be detected by fluorescence microscopy or an HCS reader. This probe is compatible with the HCS LipidTOX™ phospholipidosis detection reagents (H34350, H34351). HCS LipidTOX™ neutral lipid stains can also be used to monitor the formation and differentiation of adipocytes, a process called adipogenesis.
Confocal Microscopy Leica SP5 Inverted Confocal Microscope. Excitation frequencies used: 405, 488, 633
Single-Use RED Plate with Inserts (Thermo Scientific: 2034.6) and ICP-MS Single-Use RED Plate with Inserts (Thermo Scientific: 2034.6) followed by mass spectrometry for quantification of compound and determination of: % Free = (Concentration buffer chamber/Concentration plasma chamber) × 100% and % Bound = 100% - % Free.
DCFDA Cell permeable fluorescent and chemiluminescent probes. 2'-7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) is one of the most widely used techniques for directly measuring the redox state and ROS of a cell.
Fluorescence microscopy (ROS) Video microscopy imaging of reactive oxygen species produced in the cell via measuring the mean intensity values of DHR. Quantification of the images were performed with an in house software (JEX) developed at the Beebe lab at UW Madison.
RapidFire-MS The RapidFire 365 High-Throughput Mass Spectrometry System significantly increases the throughput of LC/MS analyses by replacing HPLC with automated solid phase extraction (SPE). An integrated robot manages reusable SPE cartridges, which are available in a variety of chemistries to optimize selectivity. The RapidFire 365 is a high speed autosampler that can be integrated with Agilent’s Triple Quadrupole, Time-of-Flight, and Quadrupole Time-of-Flight mass spectrometers. RapidFire enables automated high-capacity sample analysis in a wide range of applications, even in biological matrices. The technology provides a more successful approach to HTS of challenging drug targets and a 10x faster throughput for MS-based analyses like HT-ADME discovery assays. This end to end solution delivers shorter time to results, reduced operational costs and automated high-capacity sample processing.
Microscopy - Confocal 405, 488, 633
Biliary efflux assay Using FDA as a substrate, measuring fluroscein excretion

Studies Utilizing this Target/Analyte
Study Start Date Study Types Description
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