Vivid™ CYP450 Screening Kits are high-throughput fluorescence-based assays for detection of enzyme-drug interactions and CYP450 inhibition. They provide an optimized method for studying isozyme-specific CYP450-drug interactions, metabolism, and inhibition.
Vivid™ CYP450 Screening Kits offer:
Easy three-step procedure, 'mix and read' format, reactions at room temperature or 37°C
Superior fluorescent properties and kinetics compared to conventional fluorogenic probes
High signal-to-background ratio, broad dynamic range
Compatible with multiple assay formats from 96-well to 1536-well
Simple Mix and Read Kit Format
Vivid™ CYP450 Screening Kits include Vivid™ Substrate, Vivid™ Fluorescent Standard, reaction buffer, Vivid™ Regeneration System, NADP+, and CYP450 BACULOSOMES™ Plus Reagents. The CYP450 BACULOSOMES™ Plus Reagents are microsomes prepared from insect cells expressing a human CYP450 isozyme (CYP3A5 in this case) and human cytochrome-P450 reductase. CYP450 BACULOSOMES™ Plus Reagents offer a distinct advantage over human liver microsomes in that only one CYP450 isozyme is expressed, thereby preventing metabolism by other CYP450s.
Unique Vivid™ Reagents for Bright Fluorescent Signals and Low Background
Vivid™ Substrates are blocked dyes that yield minimal fluorescence signal until cleaved or hydroxylated. Oxidation at either of two potential sites releases the highly fluorescent product (Figure 1). They have superior fluorescence, solubility, and kinetic properties compared to conventional fluorogenic probes. This results in higher sensitivity, greater signal-to-noise ratio, and better assay reproducibility. Vivid™ DBOMF Substrate is the substrate included in this particular kit. This substrate yields a product that emits a green fluorescence.
Flexible Assay Formats for Optimized Results
The sensitivity of the Vivid™ CYP450 assays allows detection of weak inhibitors and miniaturization to as little as 2 μl per reaction. Assays may be set up in kinetic mode or in end-point mode to facilitate multi-plate screening (Figure 2). Assays may be performed at room temperature or 37°C.
Applications
High-throughput screening of enzyme-drug interactions
Compound profiling for drug inhibition of cytochrome P450 isozymes
Generation of predictive SAR models to guide compound acquisition
Vivid™ CYP3A5 Green Screening Kit, 300 Assays, Liquid, Green Color, 96-well Plate Format, Fluorescent Detection Method, CYP3A5 Detection Target, VIVID™ Green Label, Use With Microplate Reader, Used in Cell Viability, Proliferation and Function, Pharma and Biopharma, Protein Assays and Analysis, Target-Based ADME/Tox Assays
VIVID™
Protein biology, cell analysis, cell viability, proliferation and function, cell metabolism, drug discovery and development, target-based adme and frasltox assays, biochemical ADME assays, cytochrome p450 inhibition adme assays, enzyme and protein activity assays, protein assays and analysis, cytochrome p450 activity
Vivid™ CYP450 Fluorogenic substrates are also available separately.
Store reaction buffers at +4°C or room temperature. Store BACULOSOMES™, the NADPH-regeneration system, and NADP+ at -80°C. Store the Vivid™ substrates and standards at +4°C, protected from light. (For long-term storage, resuspend in organic solvent and keep at -20°C.)
VIVID Green
>300 assay(s)
Green
Microplate Reader
Liquid
Vivid™ CYP450 fluorogenic substrates, BACULOSOMES™ CYP450 reagents, fluorescent standards, and an NADPH-regeneration system
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