Intracellular Staining by Flow Cytometry
|Detection of STAT5b in Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cell line was stained with Mouse Anti-Human STAT5b Monoclonal Antibody (Catalog # MAB1584, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0101B).|
|Detection of Human STAT5b by Western Blot. Western blot shows lysates of Daudi human Burkitt's lymphoma cell line untreated (-) or treated (+) with 500 U/mL Recombinant Human IFN‑ alpha (Catalog # 11200-1) for 15 minutes. Gels were loaded with 20 μg of cytoplasmic (Cyto) and 10 μg of nuclear extracts (Nuc). PVDF membrane was probed with 1 µg/mL Mouse Anti-Human STAT5b Monoclonal Antibody (Catalog # MAB1584) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for STAT5b was detected at approximately 110 kDa (as indicated). For additional reference, duplicate lysates were probed with 1.0 µg/mL Human Phospho-STAT5a/b (Y699) Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # AF4190). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Signal Transduction and Activator of Transcription 5B (STAT5b) is a member of the Jak/STAT signal transduction pathway and is activated by a variety of cytokines (IL-22, IL-6, IFN alpha ). Upon cytokine activation, STAT5b is phosphorylated and translocates to the nucleus where it binds to the DNA consensus site TTCN3GAA and activates transcription of specific genes. STAT5b shares 93% amino acid identity with STAT5a. STAT5b is involved in a wide array of biological processes ranging from regulating apoptosis to adult mammary gland proliferation, differentiation and survival.
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