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A431 cells were either left untreated (left lane) or treated (right lane) with 100 ng/ml EGF for 5 minutes at 37°C. The top panel was probed with Stat3 (Cat. No. 610189) and the bottom was probed with Stat3 (pS727) (Cat. No. 612542). Lane 1: 1:250, lane 2: 1:500 dilution of the mouse anti-Stat3 antibody.
A431 cells were either left untreated (left lane) or treated (right lane) with 100 ng/ml EGF for 5 minutes at 37°C. The top panel was probed with Stat3 (Cat. No. 610189) and the bottom was probed with Stat3 (pS727) (Cat. No. 612542). Lane 1: 1:250, lane 2: 1:500 dilution of the mouse anti-Stat3 antibody.
A431 cells were either left untreated (left lane) or treated (right lane) with 100 ng/ml EGF for 5 minutes at 37°C. The top panel was probed with Stat3 (Cat. No. 610189) and the bottom was probed with Stat3 (pS727) (Cat. No. 612542). Lane 1: 1:250, lane 2: 1:500 dilution of the mouse anti-Stat3 antibody.
A431 cells were serum starved and treated with EGF (100 ng/ml) for 5 minutes, then fixed in 1:1 methanol-acetone for 10 minutes at -20°C. Immunofluorescent staining was performed using Stat3 (pS727) (Cat. No. 612542).
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
The 49/p-Stat3 monoclonal antibody recognizes the S727-phosphorylated form of Signal transducer and activator of transcription 3 (Stat3/STAT3 isoform 1). The fluorochrome-conjugated formats have been evaluated using a human model system. However, the unconjugated form of this antibody is also effective for western blot analysis of human, mouse, and rat tissue. The Stat proteins function both as cytoplasmic signal transducers and as activators of transcription. Seven mammalian Stat proteins have been identified: Stat1-4, Stat5a, 5b, and Stat6. Stat3 is a 92-kDa protein that is activated as a DNA binding protein through cytokines, such as IL-6, and growth factors, such as EGF. Stat3 is phophorylated at serine 727 (S727) via the MAPK pathway. The S727 residue is located at a conserved Pro-X-Ser-Pro sequence, which is recognized by the protein kinase ERK. Activation through the S727 residue is thought to lead to initiation of transcription. Upon activation, Stat3 dimerizes, translocates to the nucleus, and binds DNA response elements thereby regulating gene expression. It appears that Stat3 binds to DNA as a homodimer, but it is also capable of binding as a heterodimer with Stat1. In addition to serine phosphorylation, Stat3 is also phosphorylated at tyrosine 705 by JAK1 in response to cytokine stimulation. Stat3 is widely expressed and can bind to the sis-inducible element (SIE) site from the c-fos promoter. This site is similar to the GAS element that is present in IFN--induced genes. Thus, phosphorylation of S727 in Stat3 occurs in response to growth factors and cytokines, and is essential for normal transcriptional activity.
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.