详细信息:The Millipore MDR1 Shift Assay provides a convenient method to probe conformational changes in MDR1 that occur upon transport of MDR1 substrates. Thus, the kit may be used to determine the extent to which small molecules act as transport substrates for MDR1. Because UIC2 binding is drastically increased in the presence of MDR1 substrates, the MDR1 Shift Assay can be utilized for improved, highly sensitive and specific detection of MDR1 expression. In addition, the kit is valuable for the study of structure-function relationships in cells expressing mutant MDR1 molecules.
The kit includes the UIC2 antibody, a conformation-sensitive mouse monoclonal against human MDR1, and an isotype-matched negative control mouse IgG2a. A phycoerythrin-conjugated anti-mouse IgG is provided for sensitive detection of bound primary antibody to the cells. In addition, a potent transport substrate of MDR1, vinblastine, and the diluent control (DMSO) are provided as positive and negative controls for inducing the conformational change in MDR1. Concentrated PBS (10x) and a BSA solution are included as components for medium for washing the cells after antibody binding steps. Also included is a propidium iodide solution to stain dead cells and exclude them from analysis.
Sufficient material is provided to permit 50 tests each of control IgG2a, UIC2 with DMSO diluent control, and UIC2 with vinblastine.
Analysis with a flow cytometer is required to detect the differential immunoreactivity of MDR1 upon substrate binding with the conformation-sensitive UIC2 antibody.
