Cellomics HCS Reagent Kit for multiplex analysis of MDM2 and p53 expression. | | | | | control | treat cells | detect MDM2 and/or p53 | analyze | conclude |
DescriptionThe MDM2 and p53 Detection HCS Reagent Kits are for the simultaneous quantitation of MDM2 and p53 expression in the same cell (Figure 1). These kits allow direct measurements of MDM2 and p53 in the nucleus using a fixed end-point assay based on immunofluorescence detection in cells grown on standard high-density microplates. The primary antibodies are specific for their targets and have minimal cross-reactivity with other targets. The orange p53 and green MDM2 multiplex kits contain primary antibodies toward p53 and MDM2 rabbit polyclonal and mouse monoclonal, respectively) and secondary antibodies conjugated with DyLight 488 (green) and DyLight 549 (orange) Fluorophores. The orange MDM2 singleplex kits contain a mouse monoclonal primary antibody toward MDM2 and a DyLight 549-Conjugated Secondary Antibody. These kits have been optimized for use with the Cellomics ArrayScan HCS Reader using the Compartmental Analysis BioApplication Software Module but can be used with other Cellomics BioApplications (see the Compatible BioApplication Software Modules Section). Thus, automated plate-handling, focusing, cell image acquisition/processing and data analysis/management are combined in one high-content screening (HCS) system to assay for test compounds regulating MDM2 and p53 expression. In addition to HCS instruments, cells labeled by the reagents in this kit can be viewed and analyzed by other fluorescence microscopes. See product instructions for additional background, example data and literature references.  |  |  | Hoechst dye (nuclei) | MDM2 detection | p53 detection | Figure 1. Staining of MDM2 and p53 in A549 cells treated with 1.5 µM camptothecin for 20 hours. Except for the Hoechst nuclei control (left) non-treated cells had negligible staining of either MDM2 or p53. Proteins were detected according to the kit protocol. The cells were imaged using a Cellomics ArrayScan HCS Reader. See product instructions for further details. |
Related Poster Presentations | Bhaskar S. Mandavilli, Suk J. Hong, Krishna M. Vattem, and Richik N. Ghosh (2008) Quantitative, Cell-Based, High-Content Screening Assays for DNA Damage-Induced Cell Cycle Checkpoints. |
Kit Contents| Component | 8401801 | 8401802 | 8401901 | 8401902 | | p53 Primary Antibody | 11 µl | 55 µl | -- | -- | | MDM2 Primary Antibody | 55 µl | 275 µl | 55 µl | 275 µl | | DyLight 549 Conjugated Goat Anti-Rabbit IgG | 14 µl | 30 µl | -- | -- | | DyLight 549 Conjugated Goat Anti-Mouse IgG | -- | -- | 8 µl | 30 µl | | DyLight 488 Conjugated Goat Anti-Mouse IgG | 14 µl | 60 µl | -- | -- | | Hoechst Dye | 30 µl | 30 µl | 30 µl | 30 µl | | Wash Buffer (10X Dulbecco's PBS) | 100 ml | 100 ml | 100 ml | 100 ml | Permeabilization Buffer (10X Dulbecco's PBS with 1% Triton® X-100) | 100 ml | 100 ml | 100 ml | 100 ml | | Blocking Buffer (10X) | 85 ml | 85 ml | 85 ml | 85 ml | | Thin Plate Seal Assembly | 7/pack | 7/pack | 7/pack | 7/pack |
Ordering InformationScreening size kits and components (bulk and custom) available with special pricing. Please call. To order Cellomics HCS Reagent Kits, please call 800-874-3723 in the U.S.
or contact your local distributor of Thermo Scientific Products. Choose "Order Online" in table to see pricing and order online (U.S. only). See all Cellomics HCS Reagent Kits
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