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

The nuclear factor kB (NFkB) is a sequence-specific DNA-binding protein (2). It is a pleiotropic transcription factor which is involved in the expression of a variety of cellular and viral genes (3). NFkB consists of two subunits which are named according to their molecular weight, p50 and p65 (4). The subunits are stabilized by a so-called inhibitory chain IkB (5). In quiescent cells, NFkB resides in the cytosol in an inactive form which can be activated in vivo by treatment of cells with cytokines or protein kinase activators (6). In vitro, it is possible to generate the active form of NFkB by treatment with sodium deoxycholate, formadine or electrophoretic size fractionation. The active form of NFkB is a heterotetrameric protein, consisting of the two p50 and two p65 subunits (7). After activation NFkB translocates from the cytosol to the nucleus of the cell, binds to specific DNA sequences and initiates transcription. \n\nApplications: \nSuitable for use in Immunofluorescence, Flow Cytometry, ELISA, Western Blot, Gel Supershift, Electrophoretic mobility supershift assays (EMSA), Immunohistochemistry and Immunocytochemistry. Other applications not tested.\n\nRecommended Dilution:\nImmunofluorescence: 1-10ug/ml\nFlow cytometry: fixed cells only (acetone fixed cells)\nWestern Blot: 5-10ug/ml (ECL) \nImmunohistochemistry (Frozen, paraffin): 5-10ug/ml, APAAP\nHIER required Citrate buffer, pH6.0\nSupershift assay: 0.5-1ug/ml\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), 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. \n

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SPECIFICATIONS

Catalog Number

N2302-03

Size

100ug

Applications

ELISA, FC, ICC, IF, IHC, WB

Hosts

Mouse

Reactivities

Hum

Form

Supplied as a liquid in PBS, pH 7.6, 0.1% sodium azide.

P Type

Mab

Purity

Purified by Protein A affinity chromatography.

Isotype

IgG3

References

1. EMBO (1993) 12:201

Additional Info

Recognizes an epitope overlapping the nuclear location signal (NLS) of the p65 subunit of the NFkB heterodimer. Thus it selectively binds to the activated form of NFkB (1).