Detection of Human Phospho‑STAT3 (Y705) by Western Blot. |
Western blot shows lysates of Daudi human Burkitt's lymphoma cell line and HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 500 U/mL Recombinant Human IFN‑ alpha A (Catalog # 11100-1) for 20 minutes or 50 µg/mL Recombinant Human IL‑6 (Catalog # 206-IL) for 15 minutes. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human Phospho-STAT3 (Y705) Monoclonal Antibody (Catalog # MAB4607) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Phospho-STAT3 (Y705) at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Human STAT-3/STAT3a (signal transducer and activator of transcription 3; also called APRF) is a 90-95 kDa member of the STAT family of transcription factors. It is 770 amino acids (aa) in length, contains one SH2 domain (aa 580-670), and is found in almost all cell types. STAT3 mediates gp130, LIF-R, OB-R, IL10-R and EGFR signaling. Upon activation, receptors such as gp130 phosphorylate STAT3 at Ser727, which results in transcriptional activation. Phosphorylation at Tyr705 mediates homo- and hetero-dimerization with STAT1 and nuclear translocation, and is an important event in oncogenic transformation. The STAT3b isoform shows a six aa substitution for aa 716-770, thus eliminating the Ser727 site. Human and mouse STAT3a share 99% aa sequence identity.
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