Product Name Catalog # Price   Qty
Real-Time PCR Assay for Monitoring NFkB Regulated Genes CL-1001 $479
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Description:

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. When it is activated, NFkB mediates the expression of genes involved in immune and inflammatory responses, cell growth control, and apoptosis. To facilitate the monitoring of gene expression representing these biological functions with a limited number of cells, Signosis developed a real-time PCR assay to analyze 8 genes in total RNA or directly in cell lysates. 

Benefits:

  • All-in-One - All reagents for the experiment, from cell lysis, reverse transcription to PCR assay, are included in  one kit.
  • No need for RNA preparation - Cell lysates can be used directly in reverse transcription.
  • Robust - Full length cDNA can be obtained with the kit.
  • Real-time - Analysis can be monitored in real time.

Principle

Signosis’ Real-Time PCR assay is to monitor the expression of 8 genes through reverse transcription of mRNA into cDNA and real-time PCR. The purified total RNA or cell lysate is first reverse transcribed into cDNA.  Targeted genes are then analyzed with real-time PCR.

 

 

 

Data

 

Figure 1: Real-Time PCR Analysis of NFkB Target Genes in Cell Lysate and Total RNA. The HeLa cells were treated with TNFa for 6 hours. The cell lysate from 1000 cells and 0.1 ug of total RNA were subjected to the assay. The real-time PCR was conducted on ABI7700. 

Literature

View user manual

Citations

 1.Inositol 1,4,5-trisphosphate (IP3) receptor up-regulation in hypertension is associated with sensitization of Ca2+ release and vascular smooth muscle contractility. Abou-Saleh H, Pathan AR, Daalis A, Hubrack S, Abou-Jassoum H, Al-Naeimi H, Rusch NJ, Machaca K J Biol Chem. 2013 41958;288(46):32941-51. 

2.A long noncoding RNA contributes to neuropathic pain by silencing Kcna2 in primary afferent neurons. Zhao X, Tang Z, Zhang H, Atianjoh FE, Zhao JY, Liang L, Wang W, Guan X, Kao SC, Tiwari V, Gao YJ, Hoffman PN, Cui H, Li M, Dong X, Tao YX. Nat Neurosci. 2013 Aug;16(8):1024-31. 

3.Protein Kinase CK2 Inhibition Down Modulates the NF-κB and STAT3 Survival Pathways, Enhances the Cellular Proteotoxic Stress and Synergistically Boosts the Cytotoxic Effect of Bortezomib on Multiple Myeloma and Mantle Cell Lymphoma Cells. Manni S, Brancalion A, Mandato E, Tubi LQ, Colpo A, Pizzi M, Cappellesso R, Zaffino F, Di Maggio SA, Cabrelle A, Marino F, Zambello R, Trentin L, Adami F, Gurrieri C, Semenzato G, Piazza F. PLoS One.  2013 41544;8(9):e75280. 

4.IP3 receptor up-regulation in hypertension is associated with sensitization of Ca2+ release and vascular smooth muscle contractility. Abou-Saleh H, Pathan AR, Daalis A, Hubrack S, Abou-Jassoum H, Al-Naeimi H, Rusch NJ, Machaca K. J Biol Chem.  2013 Oct 4. [Epub ahead of print] 

5.Direct Quantification of mRNA and miRNA from Cell Lysates Using Reverse Transcription Real Time PCR: A Multidimensional Analysis of the Performance of Reagents and Workflows. Ho YK, Xu WT, Too HP. PLoS One.  2013 41522;8(9):e72463. 

6.Osteoprotegerin Contributes to the Metastatic Potential of Cells with a Dysfunctional TSC2 Tumor-Suppressor Gene. Steagall WK, Pacheco-Rodriguez G, Glasgow CG, Ikeda Y, Lin JP, Zheng G, Moss J. Am J Pathol.  2013 Sep;183(3):938-50. 

7.Rapid expression profiling of brain microvascular endothelial cells by immuno-laser capture microdissection coupled to TaqMan® Low Density Array. Demarest TG, Murugesan N, Shrestha B, Pachter JS. J Neurosci Methods., 2012 ;206(2):200-4. 

8.Nuclear Aggregation of Olfactory Receptor Genes Governs Their Monogenic Expression.Clowney EJ, Legros MA, Mosley CP, Clowney FG, Markenskoff-Papadimitriou EC, Myllys M, Barnea G, Larabell CA, Lomvardas S. Cell., 2012 41222;151(4):724-37. 

9.Active induction of experimental autoimmune encephalomyelitis by MOG35-55 peptide immunization is associated with differential responses in separate compartments of the choroid plexus. Murugesan N, Paul D, Lemire Y, Shrestha B, Ge S, Pachter JS. Fluids Barriers CNS., 2012 41125;9(1):15. 

10. Alterations in Tight Junction Protein and IgG Permeability Accompany Leukocyte Extravasation Across the Choroid Plexus During Neuroinflammation.Shrestha, Bandana BS; Paul, Debayon MS; Pachter, Joel S. PhD . Journal of neuropathology and experimental …, 2014