NFkB, p65, CT (NF-kB, Nuclear Factor kappa B, RelA)

The transcription factor NFkB is widely recognized as a critical mediator of immune and inflammatory responses. In most cell types, NFkB is found in the cytoplasm where it is associated with an inhibitory protein known as IkB. An impressive variety of stimuli (tumor necrosis factor, interleukin 1, T-cell activation signals, bacterial endotoxins, viral transforming proteins, certain growth factors and reactive oxygen intermediates) lead to the rapid nuclear accumulation of NFkB by the induced phosphorylation and subsequent degradation of IkB. In the nucleus, NFkB regulates genes encoding cytokines, cytokine receptors, cell adhesion molecules, proteins involved in coagulation and genes involved in cell growth control. NFkB is thought to be an important transcriptional regulator for HIV. Growing evidence indicates that the dysregulation of NFkB may be key to a number of diseases including arthritis and other inflammatory diseases, Alzheimer's disease, atherosclerosis and cancer. NFkB is formed through the association of multiple subunits, either as a homodimer or heterodimer. Subunits have been identified as p50 (NFkB1), p65 (RelA), c-Rel, RelB and p52 (NFkB2). The classic NFkB form exists as a p50-p65 heterodimer and predominates in many cell types. Many of the possible combinatorial forms of homo- and heterodimers have been identified. Growing evidence indicates that different forms of NFkB have different functions in cells. Interestingly, both the p50 and p52 subunits are derived from the precursor proteins p105 and p100 respectively. Each contain multiple copies of the so-called ankyrin repeat at their C-termini. Nuclear translocation of NFkB is confirmed by the use of electrophorectic mobility shift assays or by Immunoblot with nuclear extracts. The subunit composition of NFkB is confirmed by the use of antibodies that "supershift" the DNA/protein complex.\n\nApplications: \nSuitable for use Western Blot, Gel Supershift Assay, ELISA and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000-1:2500 \nGel supershift assay: 1-2ul per assay. \nOptimal dilution determined by the researcher.\n\nRecommended Reagents:\nI1904-46Q: IgG, H&L (HRP) Pab Gt xRb\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

N2302-02

Size

100ul

Applications

ELISA, IP, WB

Hosts

Rabbit

Reactivities

Hum, Rat

Form

Supplied as a liquid in PBS, 0.1% sodium azide, before the addition of glycerol to 40%.

P Type

Pab

Purity

Serum

Isotype

IgG

References

1. Unlap, M.T., Jope, R.S., Dexamethasone attenuates NF-B DNA binding activity without inducing IB levels in rat brain in vivo. Molecular Brain Research 45: 83-89 (1997). 2. Poser, I., et al Upregulation of HMG1 Leads to Melanoma Inhibitory Activity Expression in Malignant Melanoma Cells and Contributes to Their Malignancy Phenotype.Mol Cell Biol 23:8, 2991-8 (2003). 3. Jobin, C, et al., Curcumin blocks cytokine-mediated NF-kappa B activation and proinflammatory gene expression by inhibiting inhibitory factor I-kappa B kinase activity. J Immunol 163(6): 3474-83 (1999). 4. Hellerbrand, et al., Am. J. Physiology 275(2): 269-278 (1998).

Additional Info

Recognizes NFkB p65 (RelA). May react non-specifically with other proteins. Species crossreactivity: mouse (by Western Blot), mouse and rat (by Gel Shift).