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 ArrayScan® HCS Reader using the Compartmental Analysis BioApplication Software Module but can be used with other Cellomics BioApplications. 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.
Figure 2. 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.
Figures 3. Detection and analysis of MDM2 and p53 in A549 cells in response to treatment with Nutlin-3. Images are of cells treated with 6 µMnutlin-3 for 20 hours. Nuclear morphology detected with Hoechst dye (left). Detection of MDM2 (green) and p53 (orange)(right).
See product instructions for additional background, example data and literature references.
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