Akt is a serine/threonine protein kinase that plays important roles in mediating signals for numerous of cell functions including cell growth, differentiation, cell cycle, transcription, and glucose metabolism. The enzyme is activated by PI3K, and activated Akt leads to phosphorylation of a wide range of protein substrates such as GSK-3, IKK-ß, BAD, MDM2, mTOR, CASP9, p21Cip1, p27Kip1. Differential expression of Akt substrates contributes different phenotypic outcomes.
Signosis has developed a plate-based array for profiling 20+ Akt-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 sigma||CCND1||MDM2||14-3-3 sigma||CCND1||MDM2||14-3-3 sigma||CCND1||MDM2||14-3-3 sigma||CCND1||MDM2|
|D||Akt2||FOXO3||p27, kip1||Akt2||FOXO3||p27, kip1||Akt2||FOXO3||p27, kip1||Akt2||FOXO3||p27, kip1|
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 Akt pathway cDNA plate array analysis. HeLa cells were treated with and without 100ng/ml IGF for 4hours, RNAs prepared, cDNA synthesized with biotin label and subjected to cDNA plate array hybridization and detection.