A Homogeneous Multiplex Immunoassay for RTK Phosphorylation State that is Adaptable to HTS

6 Aug 2012

This application note looks at the VersaMAP™ Magnetic Human RTK Multiplex Kits which are Luminex ® bead-based multiplex immunoassays for the analysis of RTK phosphorylation. These assays can be readily adapted for high-throughput screening with minimal manual intervention from cell treatment to data collection. To demonstrate this, several different cell lines were grown in 96-well plates, treated with various stimulants or inhibitors, and lysed in the same plate.

VersaMAP™ Development System

Bio-Techne

The VersaMAP Development System offers our widest selection of potential analytes for Luminex-based multiplexing. The diluent supplied is suitable for most cell culture supernate samples. Samples with complex matrices, such as serum and plasma, may need further optimization. For multiplex kits validated by R&D Systems for use in complex matrices, please see our Fluorokine® MAP products. VersaMAP Development System product datasheet (PDF) Please use the VersaMAP Development System Configuration & Ordering Tool to select up to 40 analytes to multiplex. Features • User defines up to 40 analytes of interest • Wide selection of analytes • Requires a small sample volume (• Mass-calibrated standards for consistent results with every new lot of material • Optimized antibody pairings and concentrations saves development time • Development protocols provide directions for optimization of non-validated sample types • Enough reagents for two 96-well plates • Quick turn-around time: only 3-5 days *Please allow additional time when placing orders through a distributor. Kit Contents • Premixed antibody-coated bead cocktail • Premixed biotinylated detection antibody cocktail • Standard Cocktail(s) • Bead Diluent • Biotin Antibody Diluent • Standard/Sample Diluent • Wash Buffer • Streptavidin-PE • 2 Filter-bottom 96-well Microplates • Foil Plate Sealers • Mixing Bottle • Certificate of Analysis

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VersaMAP™ Magnetic Custom Multiplex Kits

Bio-Techne

The VersaMAP™ Magnetic Custom Multiplex Kits are our most efficient Luminex-based multiplex assays. These homogeneous, no wash assays utilize analyte-specific antibodies conjugated to superparamagnetic microparticles to simultaneously detect up to 25 user-defined phosphorylated receptor tyrosine kinases (RTKs) in a single sample. Available Kits VersaMAP Magnetic Human RTK Kit A VersaMAP Magnetic Human RTK Kit B Features • Complete ready-to-use kits • Microplate-based assay • User defines up to 25 analytes of interest • Simultaneous detection of selected analytes • Small sample size (1-5 μg of cell lysate) • Simple procedure: no washing required • Superparamagnetic beads are premixed • Compatible with multiple analyzing systems* Kit Contents • Premixed antibody-coated superparamagnetic bead cocktail • Biotinylated anti-phospho-tyrosine detection antibody • Positive Recombinant Peptide Control • Streptavidin-PE • Diluents • Lysis Buffer • 1 Round-bottomed 96-well Microplate • Foil Plate Sealers • Mixing Bottle *Compatible with Luminex MAGPIX®, Luminex 200™, and Bio-Rad® Bio-Plex® analyzers.

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A Homogeneous Multiplex Immunoassay for RTK Phosphorylation State that is Adaptable to HTS