Phospho-p53 (ser15) and p53 Detection Kit Features
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Available in multiplex or singleplex configurations
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Optimized protocol requiring no cell lysis, purification or filtration steps
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Immunofluorescence-based assay provides sensitive results without radioactivity
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Cells prepared with the HCS Reagent Kits may be analyzed directly via standard fluorescence microscopy; or HCS Imaging Systems including the ArrayScan HCS Reader
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Primary antibodies are specific for their targets and have minimal cross-reactivity with other target
These kits allow direct measurements 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 phospho-p53 and green p53 multiplex kits contain anti-phospho-p53 (Ser15) and -p53 primary antibodies (rabbit polyclonal and mouse monoclonal, respectively) and secondary antibodies conjugated with orange and green fluorophores (DyLight™ 549 Goat Anti-Rabbit and DyLight 488 Goat Anti-Mouse antibodies, respectively). The orange phospho-p53 singleplex kits contain a rabbit polyclonal primary antibody towards phospho-p53 and a DyLight 549 Conjugated Goat anti-Rabbit Secondary Antibody.
The Multiplexed and Singleplexed phospho-p53 and p53 Detection Kits have been optimized for use with Cellomics ArrayScan® HCS Reader using the Compartmental Analysis BioApplication Software Module. These kits 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 phospho-p53 and p53 detection. In addition to HCS instruments, cells labeled by the reagents in this kit can be viewed and analyzed by other fluorescence microscopy-based instruments.
Figure 1. Staining of p53 and phospho-p53 in A549 cells treated with media only (untreated) or with 5 µM camtothecin for 20 hrs (Camptothecin 5µM). Proteins were detected according to the kit protocol. The cells were imaged using a Cellomics ArrayScan HCS Reader.
Figure 2. Dose response curves using camptothecin.
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