NF-kB (nuclear factor kB) is sequestered in the cytoplasm by IkB family of inhibitory proteins that mask the nuclear localization signal of NF-kB thereby preventing translocation of NF-kB to the nucleus. External stimuli such as tumor necrosis factor or other cytokines results in phosphorylation and degradation of IkB releasing NF-kB dimers. NF-kB dimer subsequently translocates to the nucleus and activates target genes. Synthesis of IkBa is autoregulated. IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/a, IKK2/b, and IKK3/g. In vitro, IKK1/a and IKK2/b can form homo- and heterodimers that can phosphorylate IkBs at the regulatory serine residues directly. IKK1/a and IKK2/b are phosphorylated by NF-kB-inducing kinase (NIK) and MAP kinase kinase kinase-1 (MEKK1) (1), respectively. Targeted disruption of IKK1/a gene in mice results in skin and limb abnormalities and death of newborns (2,3).
Positive Control: Daudi, NIH 3T3, or Raw
Applications:
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution:
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot 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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