The NF-kB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IkB proteins (1–3). Activation occurs via phosphorylation of IkB-a at Ser32 and Ser36, resulting in the release and nuclear translocation of active NF-kB (3–7). IkB-a phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals, including inflammatory cytokines, growth factors and chemokines. Phosphorylation of IkB-a at Ser32 and Ser36 has been shown to stimulate conjugation with ubiquitin followed by proteasome-mediated degradation of IkB, resulting in the release of active NF-kB. Kinases that phosphorylate IkB at these activating sites have been identified (8). Because phosphorylation of IkB-a at Ser32 is essential for release of active NF-kB, phosphorylation at this site is an excellent marker of NF-kB activation (1–3). Nonphosphorylated IkB-a Control Cell Extracts: Total cell extracts from HeLa cells prepared without treatment serve as a negative control.
Applications:Western Blots: As controls, we recommend using 20ul of phosphorylated and nonphosphorylated IkB-a cell extracts.
Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6 months at -20°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.
仕様
Size：200ul
Source Antigen：HeLa cells
Grade：Lysate
Form：Supplied as a liquid in SDS Sample Buffer: 62.5mM Tris-HCl, pH 6.8 at 25°C, 2% w/v SDS, 10% glycerol, 50mM DTT, 0.01% w/v bromophenol blue or phenol red.
