CIKS, NT (Connection to IKK and SAPK, JNK, Act1, NF-kB Activator 1)

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 (1). External stimuli such as tumor necrosis factor or other cytokines, such as IL-1 results in phosphorylation and degradation of IkB releasing NF-kB dimers. NF-kB dimer subsequently translocates to the nucleus and activates target genes (2). Recently, Li et al. (3) have isolated a cDNA coding for a protein named Act1 (NFkB activator 1). Act1/CIKS cDNA codes for a protein containing 574aa with a predicated molecular mass of 60kD. Act1/CIKS protein contains a helix-loop-helix domain in its N-terminal portion and a coiled-coil structure at the C terminus. Act1 lacks any catalytic domain, however, it activates IKK through the helix-loop-helix domain. The activation of NF- B by Act1 appears to be through the NIK-IKK complex, because a NIK dominant-negative mutant protein inhibited Act1-induced NF- B activation. IL-1 responsive pathway is also necessary for Act1 function. Act1 mRNA is expressed in a variety of tissues, with high level in thymus, kidney and placenta; moderate level in heart, skeletal muscle, colon, liver, lung and small intestine; and at a very low level in brain, spleen, and peripheral blood leukocytes. \n\nApplications: \nPositive control: human/mouse/rat kidney whole cell lysate. Western blot analysis of CIKS using antibody on 15 ugs of mouse kidney cell lysate. Observed size: ~60 kD\n\nRecommended Dilution:\nWestern blot: 1-2ug/ml \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, aliquot and add glycerol (40-50%). Freeze 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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