NFkB-Regulated cDNA Plate Array

NFkB is a ubiquitous transcription factor that plays a key role in cellular responses to stimuli such as stress, cytokines, free radicals, ultraviolet irradiation, oxidized LDL, and bacterial or viral antigens. Incorrect regulation of NFkB has been linked to cancer, inflammatory and autoimmune diseases, septic shock, viral infection, and improper immune development. When NFkB is activated, it is dissociated from its inhibitor IkB and moves from the cytoplasm to the nucleus, where it binds to target DNA elements and positively regulates the transcription of genes involved in immune and inflammatory responses, cell growth control, and apoptosis. Signosis developed a plate-based array for profiling 20+ NFkB-regulated genes.

A. Benefits:

Based on Signosis' proprietary cDNA plate array detection technology, we developed NFkB-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. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity.