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Western blot analysis of Stat3 on a A431 cell lysate (Human epithelial carcinoma; ATCC CRL-1555). Lane 1: 1:2500, lane 2: 1:5000, lane 3: 1: 10,000 dilution of the mouse anti-Stat3 antibody.
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Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml
The Stat proteins function as both cytoplasmic signal transducers and activators of transcription. Stat91/84 (the two proteins are the result of alternate splicing-Stat91has an additional 38 C-terminal amino acids) and Stat113 were the first identified members of this protein family. With the discovery of additional members of the Stat family (Stats3 & 4), the nomenclature has been revised to indicate the Stat family members in the order of their discovery. Stat 91, 84, and 113 have become Stat1α, Stat1β, and Stat2, respectively. Stat3 is a 92 kDa protein that is activated as a DNA binding protein through tyrosine phosphorylation in response to treatment of cells with EGF and IL-6 but not with IFN-γ. Stat3 is widely expressed and can bind to DNA in the absence of Stat1α or Stat2. Stat3 can bind to the sis-inducible element (SIE) site from the c-fos promoter. The site is similar to the GAS element that is present in IFN-γ induced genes. It appears that Stat3 binds to DNA as a homodimer, but it is also capable of binding as a heterodimer with Stat1. With the isolation of Stat3 and the discovery of Stat4, it appears that the Stat family may contain many members, each with a characteristic tissue distribution and specific activating ligands.
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