Human HIF-Regulated cDNA Plate Array

Rapidly growing tumors result in hypoxic regions. Adaptive responses of most cells to hypoxia are (1) to produce VEGF and other hypoxia-induced angiogenic cytokines that promote increased tissue vascularization, thereby increasing tissue oxygenation, and (2) to switch metabolically from oxidative phosphorylation to anaerobic glycolysis. Hypoxia-inducible factor 1 (HIF-1) is an oxygen-regulated transcriptional activator that plays essential roles in the process. The HIF-1alpha subunit is oxygen-dependent ubiquitination and proteasomal degradation. In addition, cytokines including interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) stimulate HIF-1 dependent gene expression. HIF-1 increases the expression of several genes that promote blood flow and inflammation, such as vascular endothelial growth factor (VEGF). Signosis’ human HIF-regulated cDNA Plate Array allows profiling 20+ related genes.

A. Benefits:

Based on Signosis' proprietary cDNA plate array detection technology, we developed HIF-regulated cDNA plate array, which offers following benefits:

• Simple - Do the gene expression profiling as doing ELISA.

• High sensitive detection- Detected mRNA molecules are converted into multiple biotin-labeled cDNAs for sensitive detection.

• Quantitative comparison – The difference of two or more samples in gene expression can be quantitatively analyzed.

B. Principle of the technology

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. A HRP chemiluminescent substrate is added for measurement. The concentrations of cDNAs are directly proportional to the chemiluminescent intensity of the test sample.