p53 plays a crucial role in maintaining ordered proliferation, growth, and differentiation of normal cells. While p53 normally exists in the cell at a very low level and in a relatively inactive form, in response to cellular stress it accumulates in the nucleus, where it binds to specific DNA sequences. p53 is known to induce or inhibit the expression of more than 100 target genes. Mutations of the p53 gene is the most frequently encountered genetic event in cancer, thereby inactivating p53 and resulting in the disruption of the expression of target genes. Therefore, expression profiling of p53 target genes will facilitate the functional study of p53.
Signosis has developed a plate-based array for profiling 20+ p53-regulated genes. By using this assay, researchers are able to compare gene expression in up to three samples on a single microtiter plate.
List of Applicable Genes
|A||14-3-3||Cyclin B||MSH2||14-3-3||Cyclin B||MSH2||14-3-3||Cyclin B||MSH2||14-3-3||Cyclin B||MSH2|
|F||cdc25||MDM2||TGF beta||cdc25||MDM2||TGF beta||cdc25||MDM2||TGF beta||cdc25||MDM2||TGF beta|
Signosis’ proprietary cDNA plate array is a plate-based hybridization profiling analysis for monitoring the expression of dozens of genes through reverse transcription of mRNA into cDNA. Like array analyses, total RNA is first reverse transcribed into cDNA in the presence of biotin-dUTP in the assay. Targeted genes are then specifically captured onto individual wells on a plate, instead of membranes, through a pre-coated gene-specific oligonucleotide. The captured cDNAs are further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity.
Human p53-regulated cDNA plate array analysis. Total RNAs from 293 and Hela were prepared. RNAs were subjected to p53-regulated cDNA plate assay, and detected with chemiluminence by a plate reader
LiteratureView user manual
Nitration of the Tumor Suppressor Protein p53 at Tyrosine 327 Promotes p53 Oligomerization and Activation. Yakovlev VA, Bayden AS, Graves PR, Kellogg GE, Mikkelsen RB. Biochemistry., 2010 Jun 29;49(25):5331-9. doi: 10.1021/bi100564w.