NF-kappa B, NF-{kappa}B, NF-kB, Nuclear Factor-Kappa B, Activation Assay Kit (Part No. NFKB-1)

NF-kappa B, (NF-{kappa}B, NF-KB) Activation Assay Kit 

Part NFKB-1;  Kit without antibodies to p65.  View kit Part No NFKB-2 if you require matched primary and secondary antibodies to detect p65.

Highlights the NF-kappa B Activation Assay Kit, Part No. NFKB-1

• Rapid assay to detect and quantitate NF-kappa B activation by isolating nuclear and cytoplasmic fractions.
  
• Nuclear and cytoplasmic fractions are isolated in less than 1 hour using a microcentrifuge.
• The NF-KB Activation Assay Kit is applicable in virtually all mammalian cells.
• For 50 to 150 NF-kappa B activation assays based on one assay per semi-confluent 10 cm cell culture dish or well of a 6-well dish, respectively.
  

 View Protocol

Figure Legends:  Representative data derived from the FIVEphoton Biochemicals NF-kappa B Activation Assay Kit in transfected HEK293 cells:  Fig. 1.  Western blot and quantitative analysis revealing NF-kappa B activation in three experimental repetitions elicited by transfection and overexpression in HEK293 cells of SNP variants of a gene associated with a neurological disease:  SNP overexpression activates a cellular stress response in HEK293 cells.  Lane 1 in the Western blot and corresponding bar chart reflect the mock transfection control (nl).  Lane 2 and the corresponding bar chart display the NF-kappa B response to expression of mutant variant SNP-1 (S1) of the gene.  Lane 3 in the Western blot and corresponding bar chart reveal the NF-kappa B response due to expression of SNP-2 (S2). Fig. 2:  Data from an independent experiment revealing elevated translocation of p65 into the nuclear fraction.  Fig. 3:  Activation of NF-kappa B and increased levels of p65 protein in the nuclear fraction following TNFalpha treatment of HEK293 cells. TNFalpha activates the NF-kappa B pathway.

NF-kappa B Activation Assay Kit Components

1. Cytoplasmic Fractionation Reagent  (CER-1;  55 ml)
2. Nuclear Fractionation Reagent (NER-1;  3.5 ml)
3. DTT (sufficient to make a IM solution, volume 64 µl)

Related NF-KB Activation Assay Kits

1.   Chen L, Yang G, Zhang X, Wu J, Gu Q, et al. 2009. Induction of MIF expression by oxidized LDL via activation of NF-kappaB in vascular smooth muscle   cells. Atherosclerosis 207: 428-33.Hoffmann, A. and Baltimore, D. (2006) Circuitry of nuclear factor kappaB signaling. Immunol Rev 210, 171-186.   
2. Hoffmann F, .Sass G, Zillies J, Zahler S, Tiegs G, et al. 2009. A novel technique for selective NF-kappaB inhibition in Kupffer cells: contrary effects in fulminan hepatitis and ischaemia-reperfusion. Gut 58: 1670-8.
3. Itoh M, Fu L, Tohda S. 2009. NF-kappaB activation induced by Notch ligand stimulation in acute myeloid leukemia cells. Oncol Rep 22: 631-4.
4. Karin, M. (2006) Nuclear factor-kappaB in cancer development and progression. Nature 441 (7092), 431-436.
5. Karin, M. and Greten, F.R. (2005) NF-kappaB: linking inflammation and immunity to cancer development and progression. Nat Rev Immunol 5 (10), 749-759.
6. Osterlund C, Gronlund H, Polovic N, Sundstrom S, Gafvelin G, Bucht A. 2009. The non-proteolytic house dust mite allergen Der p 2 induce NF-kappaB and MAPK dependent activation of bronchial epithelial cells. Clin Exp Allergy 39: 1199-208.
7. Wirth, T. and Baltimore, D. (1988) Nuclear factor NF-kappa B can interact functionally with its cognate binding site to provide lymphoid-specific promoter function. Embo J 7 (10), 3109-3113.
8. Wu S, Yin R, Ernest R, Li Y, Zhelyabovska O, et al. 2009. Liver X receptors are negative regulators of cardiac hypertrophy via suppressing NF-kappaB   signalling. Cardiovasc Res 84: 119-26.
9. Zhang L, Altuwaijri S, Deng F, Chen L, Lal P, et al. 2009. NF-kappaB regulates androgen receptor expression and prostate cancer growth. Am J Pathol 175: 489-99