Vivid™ CYP3A5 Green Screening Kit

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