PREDIVEZ Vesicular Transport Kits

Introduction

The PREDIVEZ Vesicular Transport kit is a simple and powerful tool to investigate drug transporter interactions. It provides information on any interaction between the ABC transporter and the drug candidate that would affect the transport of the fluorescent or radiolabeled probe substrate.

Assay Principle

 "Inside-out" membrane vesicles containing ABC transporters are applied in the vesicular transport assay to determine the IC50 of the compound. After incubating the test article with the inverted membranes, the membranes are then separated from the drug-containing buffer and washed before measuring their contents for transported compound (fluorescence or liquid scintillation counting).

Indirect inhibitory assay

The standard vesicular transport assay is an indirect inhibitory assay, performed with cold test articles. This assay provides information on any interaction between the ABC transporter and the test article. The transport of the reporter substrate is measured in the presence of the test article (typically in 7 concentrations) and IC50 is defined as the concentration inhibiting the transport of the reporter substrate by 50%.

Direct assay

Should radiolabeled form of the investigated compound or adequate analytical methods (LC/MS, HPLC) be available, the vesicular transport assay may be performed in a direct format without the reporter substrate and may identify substrate nature of the test article. Direct vesicular transport assay is low throughput assay. It is suitable for low permeability test compounds as high permeability compounds may escape from the vesicles through the lipid bilayer. Standard assays are always recommended before running the direct measurements with the test compound.

Why using PREDIVEZ?

Features

• The reporter substrate provides low non-specific binding, so you can get a larger signal and smaller standard deviations for your test compounds.
• Sensitive and quick fluorescent measurement With the use of a fluorescent reporter substrate you can avoid radioactive materials and lengthy scintillation counting.
• Larger dynamic range / signal-to-background ratio compared to the radioactive Vesicular transport assay
• Instant results. Data can be presented in transport values (with the help of the calibration curve, included), or in percentages. Negative control vesicles are also included in the kit for performance monitoring.

Reproducible results

• Low inter-assay variability
• Z' factor is ~ 0.74
• Increased automated system compatibility

Which PREDIVEZ for me?

PREDIVEZ vesicular transport kits are available in three sizes 3, 6 and 9 compound sizes. Kits contain sufficient amount of reagents and membrane preparation for testing 3 compounds per 96-well plate. Each kit is accompanied by a CD that includes the assay protocol, Excel spreadsheet with data processing template, MSDS and inhibition curves with reference compounds.

PREDIVEZ Fluorescent Vesicular Kits are available for all relevant membrane transporters from SOLVO's portfolio.

PREDIVEZ Radiolabeled Vesicular Kits
The Human & Rat MDR1 kits don't include reference compound. The ³H-N-methyl-quinidine is recommended but not included.

SB-MDR1/P-gp-PREDIVEZ-VT
SB-ratMdr1b-PREDIVEZ-VT