Introduction

Transporter proteins are integral components of cellular membranes, mediating the movement of molecules, including drugs, across cell barriers. Within these families, the ATP-binding cassette (ABC) transporters and solute carrier (SLC) transporters play pivotal roles in drug absorption, distribution, metabolism, and excretion (ADME). An exemplary member of the ABC transporter family, Multidrug Resistance Protein 1 (MRP1), significantly influences drug pharmacokinetics and resistance. At Creative Biolabs, we offer a sophisticated human transporter MRP1 fluorescence-based antagonist assay service to evaluate potential drug-transporter interactions.

Our Human Transporter MRP1 Fluorescence-based Antagonist Assay Service

Creative Biolabs provides a robust fluorescence-based antagonist assay to scrutinize the interaction of novel molecular entities (NMEs) with MRP1, encoded by the gene ABCC1. This protein is essential in the efflux of drugs and other xenobiotics out of the cells, affecting their intracellular concentrations. The assay utilizes HEK293 cell lines expressing recombinant full-length human MRP1, monitoring the transporter's activity via the detection of Calcein-AM, a fluorescence substrate.

Service Details

The Human Transporter MRP1 Fluorescence-based Antagonist Assay by Creative Biolabs employs a functional, cell-based approach that leverages the spectrophotometric detection of fluorescent signals. The assay protocol includes incubation of HEK293 cells-expressing MRP1 with Calcein-AM and the test compounds, under specified conditions, followed by measurement of fluorescence intensity. A decrease in fluorescence indicates effective antagonism of MRP1 transporter activity.

Assay Parameters

• Host Cell Type: HEK293
• Transfected Protein: Full-length human MRP1
• Substrate: Calcein-AM
• Incubation Conditions: 35 minutes at 37°C
• Detection Method: Spectrophotometric measurement of fluorescence
• Criteria for Significance: 25% inhibition
• Test Concentration/Dose: Dose-response study recommended with 8 concentrations in 2 replicates per concentration

Applications of Human Transporter MRP1 Fluorescence-based Antagonist Assay

Understanding how drugs interact with MRP1 is crucial in predicting and improving their pharmacokinetic profiles. The assay helps in evaluating the efflux capacity of MRP1, aiding in drug design to overcome resistance. It is employed in:

• Drug Development: Determining the impact of potential drugs on MRP1 activity, aiding in the selection of candidates with optimal ADME profiles.
• Pharmacokinetic Studies: Evaluating the role of MRP1 in drug absorption, distribution, and excretion.
• Safety Profiling: Identifying potential drug-drug interactions and toxicities related to MRP1 modulation.

Features of Our Services

Frequently Asked Questions

Q1: How does the MRP1 antagonist assay improve drug discovery?

A1: By identifying interactions early in development, the assay facilitates the selection of compounds with favorable ADME and safety profiles, potentially reducing the time and cost associated with drug development.

Q2: What makes Calcein-AM suitable for this assay?

A2: Calcein-AM is non-fluorescent until hydrolyzed within viable cells, where it emits a strong signal. The detectable reduction in fluorescence in the presence of MRP1 antagonists makes it an ideal substrate for this assay.

Q3: What are the substrate and inhibitors used in this assay

A3: The assay uses Calcein-AM as the fluorescent substrate. Test sample requirements vary depending on whether a screening or dose-response study is conducted, necessitating minimum volumes and pre-weighed amounts of the test compounds. This enables a precise evaluation of the interaction profiles of new drug candidates with MRP1.

Creative Biolabs is committed to advancing drug discovery with state-of-the-art assays and unparalleled scientific expertise. Our Human Transporter MRP1 Fluorescence-based Antagonist Assay Service is designed to support your endeavors in creating safer, more effective therapeutics.

For Research Use Only | Not For Clinical Use