Signal Transducer and Activator of Transcription 3 (Stat3) is a transcription factor that mediates the nuclear action of many different cytokines and growth factors. It plays roles in cell growth and apoptosis. Stat3 also participates in cellular transformation and tumorigenesis. The aberrant activation of Stat3 in tumor cells has been considered as a drug target. Stat3 is activated by tyrosine phosphorylation in response to a wide range of cytokines, growth factors, and hormones. Tyrosine phosphorylation triggers dimerization and nuclear translocation of the protein. Stat3 mediates the expression of its target genes that are involved in a diverse array of biological processes such as oncogenesis, cell growth, and differentiation.
Signosis has developed a plate-based array for profiling 20+ Stat3-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
|C||b-actin||cyclin E||Mcl-1||b-actin||cyclin E||Mcl-1||b-actin||cyclin E||Mcl-1||b-actin||cyclin E||Mcl-1|
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 Stat3-regulated cDNA plate array analysis. HeLa cells were treated with and without 100ng/ml Oncostatin M for 1 hours, RNAs prepared, cDNA synthesized with biotin label and subjected to cDNA plate array hybridization and detection.