|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‑22 (Catalog # 782-IL) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-STAT3 (Y705) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4607) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for STAT3 at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Detection of Phospho-STAT3 (Y705) in IFN-alpha-treated Human Daudi Cell Line by Flow Cytometry. Daudi human Burkitt's lymphoma cell line was unstimulated (light orange filled histogram) or treated with 500 U/mL rhIFN-alpha for 20 minutes (dark orange filled histogram) was stained with Human Phospho-STAT3 (Y705) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4607) or control antibody (Catalog # AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.|
|STAT3 in Daudi Human Burkitt's Lymphoma Cells. Phospho-STAT3 was detected in immersion fixed IFN-alpha treated Daudi human Burkitt's lymphoma cell line using 10 µg/mL Human Phospho-STAT3 (Y705) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4607) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
Signal Transducer and Activator of Transcription (STAT) proteins are transcription factors activated in response to cytokine, growth factor, or hormone receptor signaling. Janus kinases (JAKs) phosphorylate STAT proteins and induce dimerization. Homo- or heterodimers translocate to the nucleus where they bind to DNA and activate transcription.
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