SPECIALIZED EQUIPMENT AVAILABLE:
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High Content Imaging
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IN Cell Analyzer 6000 (GE Healthcare)
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Functionalities:
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Time-lapse
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Temperature, CO2 and humidity controls
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Confocal option
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Time-lapse microscope
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IX83 (Olympus)
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Functionalities:
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Brightfield, fluorescence (TRITC/FITC/DAPI)
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Time-lapse; motorized stage
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Temperature, CO2 and humidity controls
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Z-stack option
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Multimode Readers
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FluoSTAR Omega (BMG Labtech)
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Functionalities:
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Absorbance, Fluorescence, Luminescence detection modes
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Bottom optics and multi-mode shaking
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Built-in reagent injectors available
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CLARIOstar (BMG Labtech)
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Functionalities:
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Absorbance, fluorescence, luminescence, polarized fluorescence, time resolved fluorescence and alpha-screen (Amplified Luminescent Proximity Homogeneous Assay) detection modes
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Equipped with LVF MonochromatorTM technology (Linear Variable filters) for greater sensitivity of fluorescence and luminescence readings with adjustable bandwidths up to100 nm
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Top and bottom optics; Built-in reagent injectors available
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Flow Cytometer
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Attune NxT Flow Cytometer (Thermo Fisher Scientific)
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Functionalities:
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Multi-parametric single cell analysis with minimal compensation panels
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Rapid processing and detection of multiple samples and of rare events
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FSC, SSC, 14 fluorescent colors with PMTs, and user changeable detection methods are available for the following filters: Blue: 488 nm (50 mW), Violet: 405 nm (50 mW), Red:637 nm (100 mW) Yellow: 561 nm (50 mW)
Quantitative PCR
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QuantStudio5 (Thermo Fisher Scientific)
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Functionality:
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Equipped with 96-well thermal blocks with 0.1mL well volume that can accommodate 3 independent temperature zones for PCR optimization (VeriFlex technology)
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Runs can be monitored remotely through the Thermo Fisher Cloud
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PCR Machines
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C1000 Touch Thermal Cycler (Bio-Rad)
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Functionality:
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Dual block PCR machine capable of running 2 independent protocols simultaneously, with 48 PCR reactions in 0.2mL tubes per block
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Equipped with gradient capability to accommodate 6 temperature zones
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PCRmax Alpha Cycler 4 (Cole Parmer)
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Functionality:
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Quad block PCR machine capable of running 4 independent protocols simultaneously, with 96 PCR reactions in 0.2mL tubes per block
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Equipped with gradient capability to accommodate at least 12 temperature zones
Nucleofector/Electroporator
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Neon Transfection System (Thermo Fisher Scientific)
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Functionality:
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Specializes in highly efficient transfection of primary cells, stem cells and difficult-to-transfect cells
Chemidoc System
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Chemidoc Touch Imaging System (Bio-Rad)
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Functionality:
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Capable of visualizing nucleic acid gels (Ethidium bromide, Gel Red, Sybr GREEN), protein gels, and ..
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western blots (Chemiluminescence, colorimetric, alexa 488, etc)
Automated cell counter
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TC20 (Bio-Rad)
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Countess II FL Automated Cell Counter (Thermo Fisher Scientific)
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Benchtop Lyophilizer
DMBEL Laboratory Services
General Overview
The Laboratory Services arm of the Disease Molecular Biology and Epigenetics Laboratory (DMBEL) offers a suite of mostly cell-based assays to serve the needs of drug discovery projects and third-party clients. Bioactive hits from primary screens can be tested for drug-induced hepato-, nephro- and cardiotoxicity, which are responsible for high attrition rates in early-stage drug discovery. Confirmatory orthogonal and secondary assays are available to eliminate artefactual drug bioactivities from primary screens due to pan-assay interference compounds and color effects. Most assays are in 96- or 384-well format and some are pathway-specific, to provide clues on mechanism of action or the signaling cascade in which an extract/ compound impinges. High-content imaging is used in many of the assays, allowing physiologically relevant readouts that are able to visualize subcellular and organelle phenomena. The medium-to-high throughput nature of HCI also affords multi-parametric information from each sample and from individual cells, delivering insights on a compound’s mode of action and/or non-specific effects early on.
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ASSAYS AVAILABLE:
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IN VITRO TOXICITY ASSAYS
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Hepatotoxicity Testing via Lactate Dehydrogenase Quantification
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Nephrotoxicity Testing via Lactate Dehydrogenase Quantification
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Cardiotoxicity Testing via Lactate Dehydrogenase Quantification
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Fluorescence-based Cardiotoxicity Testing via Potassium Ion Channel Assay
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CONFIRMATORY
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Inflammation
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Luminescence-based Assessment of COX-2 Transcriptional Activity
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High-Content Imaging Fluorescence-based Detection of 5-LOX
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High-Content Imaging-based Detection and Quantification of Reactive Oxygen Species by DCFDA Staining
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High-Content Imaging-based Immunocytochemical Detection and Quantification of Prostaglandin E2
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Expression Profile Analysis of Inflammation Markers by RT-qPCR
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Diabetes
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Luminescence-Based Glucose Uptake–Glo Assay
Hypercholesterolemia
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High-Content Imaging-based Fluorescence Detection of Cholesterol Uptake via Filipin Cytochemical Staining
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High-Content Imaging-based Fluorescence Detection of LDL Uptake via BODIPY FL LDL Live Cell Staining
Hypertension
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Determination of ACE Activity using Abz-FRK(Dnp)P-OH Substrate
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Gout
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Fluorescence-based Xanthine Oxidase Inhibition Assay
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Fluorescence-based Uric Acid Quantification Assay
Obesity
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Colorimetric Quantification of Lipid Droplet Accumulation via Oil Red O Staining
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Colorimetric and Fluorometric Quantification of Triglyceride Content
Cancer: Cell Proliferation
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Luminescence-based Detection of ELK Pathway Activation
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RealTime-Glo Cell Viability Assay
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Vybrant® MTT Cell Proliferation Assay
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Cell Titer 96® Aqueous One Solution MTS Cell Proliferation Assay
Cancer: Apoptosis
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Luminescence-based Detection of Caspase 3/7 Activity
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Luminescence-based Detection of Caspase 8 Activity
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Luminescence-based Detection of Early Apoptosis using the RealTime-Glo Annexin V Apoptosis and Necrosis Assay
Cancer: Migration and Invasion
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Scratch Wound Healing Assay
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Assessment of Tumor Cell Migratory Capacity via Boyden Chamber Assay
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Expression profile analysis of EMT Markers by RT-qPCR
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Expression profiling of EMT Markers by Western Blot analysis
Cancer: Angiogenesis
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High-Content Imaging-Assisted Vessel Formation Assay
Cancer: Cytoskeletal Organization
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Phalloidin Staining of F-actin Filaments
High-Content Imaging
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Image Acquisition using IN Cell Analyzer 6000 High-Content Imaging System​
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EMAIL US AT:
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dmbel.labservices.upd@up.edu.ph
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