MEDINA develops different phenotypic and target-based assays, both on-demand and optimized tests. Custom development from your tube assay to a microtiter plate (96/384) assay for High-Throughput Screening (HTS), hit confirmation, hit-to-lead and Mechanism of Action (MoA) of the lead.
Cell-Based Assays
- Kinetics and endpoint measurements
- Cell viability: MTT, Resazurin, Neutral Red Uptake (NRU), ATP level, LDH, propidium iodide nuclear staining. Cell line panel > 50 cell lines
- Ion channels: hERG, Cav1.2 and Nav1.5 (cardiotoxicity)
- GPCRs: serotonin and dopamine receptors (neurobiology)
- Protection of reactive oxygen species (ROS) formation (neurobiology & antitumor)
- Nitric oxide levels and interleukins measurement (TNF-α/Il-1β/IL-6/IL-8) in cell culture of macrophages and in intestinal epithelial Caco-2 cells (immunomodulation)
- Mitochondrial potential measurement (oxidative stress)
- Caspases (apoptosis)
- Cell cycle (apoptosis)
- Nuclear translocation: FOXO (transcription factor) and NES (Nuclear Export signal to translocate through pore complex), targets available for cellular High Content Screening (antitumor)
- Cholesterol measurement (anticholesterolemic)
- Spheroids-3D cell culture: image-based screening (HCS) & cytotoxicity (CellTiter GLO®) (antitumor)
- On-demand assay development for your protein applications (see Protein Lab section):
- Expression profiling: RT-PCR, western blot, SDS-PAGE.
- Bimolecular Complementation assays, for the functional validation of transient Protein-Protein interactions and novel competitive inhibitors.
- Bioluminescence Resonance Energy Transfer assays, reporting for the activity of your target of interest and novel inhibitors.
- Cellular compartmentalization of your target of interest in the High Content Screening format. Characterization of novel therapeutic relocators.
- Cell-painting, for bio-profiling purposes.
- Absorbance
- Fluorescence: FI, FRET, TRF-FRET (HTRF®), FP, Bimolecular Complementation (BiMC),
- Luminiscence: glow, flash, dual luminescence, BRET, NanoBRETPM
- AlphaScreen®, AlphaLISA®
- LC-MS/MS; LC-HRMS
- SelecT robot, automated cell culture system
- Liquid Handling Systems (from 1 µl to 1 ml) with 96- and 384-pipetting heads (Freedom EVO, Tecan; Biomek FX e i7, Beckman; EP3, PerkinElmer)
- Echo® Acoustic Liquid Handling System (from 15 nl to 1 µl), Beckman
- Multidrop Combi System (from 0.5 µl to 2.5 ml) for 96- and 384-well plates (ThermoFisher)
- Automatic cell counter (Countess, Invitrogen; CEDEX, Roche innovates)
- Multiplate reading system (Envision & Victor-2, PerkinElmer)
- High Content Screening System (Operetta CLS, PerkinElmer; BD Pathway 855, Becton Dickinson)
- FLIPR Tetra® High-Throughput Cellular Screening System (Molecular Devices)
- Flow cytometry (BD Accuri C6, Becton Dickinson)
- Leica AF 6000 Confocal Microscope; Zeiss Axiovert Inverted Microscope
- Odyssey Infrared Imaging System (Li-COR Biosciences)
- CFX96/384 Real-Time PCR Detection System (Biorad)
- Mass spectrometry (Api4000 (Sciex) and QTOF5600 (Sciex))
- HPLC-DAD (Agilent 1290 DAD)
Quality control: We perform QC of each screening plate automatically on signal/background ratio and Z´ score calculations, considering the best plate layout for your assay; a statistically optimal number of repeats are performed for the internal positive and negative controls, and/or a dose-response curve of relevant pharmacological standards when available. Genedata Screener® software and in-house applications help us perform the analysis, visualizations, and management of screening data.
