p53 and p21 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 targets
These kits allow direct measurements of p53 and p21 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 p21 multiplex kits contain primary antibodies towards p53 and p21 and DyLight™ 549 Goat Anti-Rabbit and DyLight 488 Goat Anti-Mouse Secondary Antibodies. The orange p53 singleplex kits contain a rabbit polyclonal primary antibody towards p53 and a DyLight 549 Conjugated Goat Anti-Rabbit Secondary Antibody. The orange p21 singleplex kits contain a mouse monoclonal primary antibody towards p21 and a DyLight 549 Conjugated Goat Anti-Mouse Secondary Antibody.
The Multiplexed and Singleplexed p53 and p21 Detection Kits have been optimized for use with the Thermo Scientific 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 p53 and p21 expression. 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 p21 and p53 in A549 cells treated with media only (untreated) or with 5 μM Camtothecin for 20 hrs. Proteins were labeled according to the kit protocol. The cells were imaged using a Cellomics ArrayScan HCS Reader.
Figure 2. Dose Response curves using Camptothecin for p53 and p21 detection.
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