The IKK Isoform Antibody Sampler Kit provides an economical means to investigate NFkB signaling within the cell. The kit contains primary and secondary antibodies to perform four Western mini-blots with each antibody.
The NFkB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IkB proteins (1-3). Most agents that activate NFkB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degradation of IkB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IkB kinase (IKK) complex, whose catalysis is generally carried out by three tightly associated IKK subunits. IKKa and IKKb serve as the catalytic subunits of the kinase. IKKg serves as the regulatory subunit (8,9). Activation of IKK depends on phosphorylation; Ser177 and Ser181 in the activation loop of IKKb (176 and 180 in IKKa) are the specific sites whose phosphorylation causes conformational changes resulting in kinase activation (10-13).
Applications:
Suitable for use in Western Blot. Other applications not tested.
Recommended Dilution:
Western Blot: 1:1000
Optimal dilutions to be determined by the researcher.
Storage and Stability:
For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 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.