NFkB is a ubiquitous transcription factor that plays a key role in cellular responses to numerous stimuli such as stress, cytokines, ultraviolet irradiation, and bacterial or viral antigens. Incorrect regulation of NFkB has been linked to cancer, inflammatory and autoimmune diseases, and septic shock. When NFkB is activated, it dissociates from its inhibitor IkB and moves from the cytoplasm to the nucleus. Once in the nucleus, 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 has developed a plate-based array for profiling 20+ NFkB-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
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 NFkB-regulated cDNA plate array analysis. HeLa cells were treated with (red bars) or without (light blue) 10ng/ml TNF for 30 min. Cells were collected for profiling 23 genes with the cDNA plate array assay (top). Time course of IL-6 induction to TNFa treatment in HeLa cells was monitored with the cDNA plate array assay.
LiteratureView user manual
1. Vascular cell-adhesion molecule-1 plays a central role in the proangiogenic effects of oxidative stress. Dong A, Shen J, Zeng M, Campochiaro PA. Proc Natl Acad Sci U S A. 2011 Aug 30;108(35):14614-9.
2. TRIM21, a negative modulator of LFG in breast carcinoma MDA-MB-231 cells in vitro. J Müller, V Maurer, K Reimers, PM Vogt, V Bucan. International Journal of Oncology .Published online on: Wednesday, September 16, 2015
3. Role and regulation of p65/b-catenin association during liver injury and regeneration: a ‘complex’ relationship. Kari Nejak-Bowen, Akshata Moghe, Pamela Cornuet, Morgan Preziosi, Shanmugam Nagarajan, Satdarshan PS Monga. GE-2016-0020 Gene Expression: The Journal of Basic Liver Research early e-pub; provisional acceptance Apr. 27, 2017. DOI: 10.3727/105221617X695762.