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- BD Rhapsody™ TCR/BCR Profiling Assays
- BD® OMICS-Guard Sample Preservation Buffer
- BD Rhapsody™ ATAC-Seq Assays
- BD Rhapsody™ TCR/BCR Next Multiomic Assays
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Western blot analysis for Stat1 (pY701) (far left figure). A431 cells (Human epithelial carcinoma; ATCC CRL-1555) were either left untreated (lane 1) or treated with 100 ng/ml EGF for 5 minutes at 37°C (lane 2). The top panel was probed with a mouse anti-Stat1 antibody (Cat. No. 610115) while the bottom panel was probed with the mouse anti-Stat1 (pY701) antibody at a 1:1000 dilution. Immunofluorescence staining for Stat1 (pY701) (middle left figure). A431 cells (Human epithelial carcinoma; ATCC CRL-1555) were either untreated (top left and bottom left quadrants) or were serum starved and then treated with 100 ng/ml EGF for 5 minutes, then fixed in 3.75% paraformaldehyde with 0.2% Trtion-X 100 (top right and bottom right quandrants). Immunofluorescent staining was performed with a mouse anti-Stat1 antibody (Cat. No. 610115) (top left and top right quadrants) and the mouse anti-Stat1 (pY701) antibody (bottom left and bottom right quadrants). Flow cytometric staining for Stat1 (pY701) (middle right and far right figures). U-937 cells (Human histiocytic lymphoma; ATCC CRL-1593.2) were either untreated (unshaded histograms) or serum starved overnight and treated with 1000 units/mL of IFN-γ for 15 min (shaded histograms). Cells were fixed with 1% formaldehyde, followed by 80% ethanol and BD Cytofix/Cytoperm™ (Cat. No. 554714). Cells were then stained with a mouse anti-Stat1 antibody (Cat. No. 610185) (middle right figure) or the mouse anti-Stat1 (pY701) antibody (far right figure).
BD Transduction Laboratories™ Purified Mouse Anti-Stat1 (pY701)
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Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml.
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Stat (Signal transducer and activators of transcription) proteins are critical mediators of the biologic activity of cytokines, including interleukins, interferons, erythropoietin, and growth factors. Ligand-receptor interaction leads to activation of constitutively associated JAK family kinases and subsequent recruitment/activation of Stat proteins by tyrosine phosphorylation. Active Stat proteins then move to the nucleus to promote transcription of cytokine-inducible genes. Seven Stat proteins have been cloned, each of which is differentially expressed and/or activated in a cytokine-specific and cell type-specific manner. Stat1 and Stat2 are components of the ISGF3 (Interferon-Stimulated Gene Factor 3) complex, which is the primary transcription activator induced by the binding of the interferon to a specific cell-surface receptor. Stat1 has two alternatively spliced isoforms, 91-kDa Stat1α and 84-kDa Stat1β; Stat1α has 38 additional C-terminal amino acids. In response to the binding of IFNα, IFNγ, EGF, PDGF, or CSF-1 to their respective receptors, the Stat1 subunits become tyrosine-phosphorylated at Y701, and the complex is translocated to the nucleus. This results in the formation of an active complex that includes the DNA-binding p48 subunit. This complex is responsible for modulating the transcription of the interferon-stimulated genes (ISGs).
The 14/P-STAT1monoclonal antibody recognizes the phosphorylated Y701 in Stat1α and Stat1β.
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