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

Manufacturer Fivephoton
Category Products / Kits
Type Kit
Specific against other
Amount 1 kit
ArtNr NFKB-1
Eclass 6.1 32161000
Eclass 9.0 32161090
Available
Highlights
Rapid assay to detect and quantitate NF-kappa B activation by isolating nuclear and cytoplasmic fractions and detection of NF-KB transcription factor subunit (i.e. p65 and p50) enrichment in the nuclear fraction by Western blot.
Nuclear and cytoplasmic fractions are isolated in less than 1 hour using a microcentrifuge.
Data from this NF-KB Activation Assay Kit are higher resolved, easier to quantitate and more convincing than fluorescence and luminescence assays, including ELISA.
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.
Background
NF-kappa B Activation Assay Kit:
The NF-kappa B Activation Assay Kit contains pre-configured reagents to assay NF-kappa B activation by separating cells into nuclear and cytoplasmic fractions, and by detecting transcription factor enrichment in the nuclear fraction. Cell fractionation into cytoplasmic and nuclear fractions is conveniently performed in 1.5 ml tubes and a microcentrifuge in approximately 45 minutes. Nuclear proteins, including activated transcription factor subunits of the NF-kappa B complex, such as p65, are enriched in the nuclear fraction. The transcription factor subunits of the NF-kappa B complex in the nuclear and cytoplasmic fractions are detected and quantitated with Western blot analysis.
NF-kappa B Activation Assay:
To separate cytoplasmic from nuclear fraction following NF-kappa B activation, cells are first exposed to the "Cytoplasmic Fractionation Reagent." The cell suspension is briefly centrifuged in a microcentrifuge, which generates the cytoplasmic fraction in the supernatant. The pellet is then washed in the Cytoplasmic Fractionation Reagent and extracted with the "Nuclear Fractionation Reagent." Aliquots of each fraction are resolved in Western blots with consecutive lanes loaded to display total cell lysate, cytoplasmic fraction and nuclear fraction. The extent of translocation from the cytoplasm to nucleus reveals a quantitative assessment of NF-kappa B activation.
References
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.

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.

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.

Karin, M. (2006) Nuclear factor-kappaB in cancer development and progression. Nature 441 (7092), 431-436.

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.

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.

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.

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.

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
Amount: 1 kit
Available: In stock
Listprice: €231.75
Price: €231.75
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