Detection of STAT6 in Daudi Human Cell Line by Flow Cytometry.
Daudi human Burkitt's lymphoma cell line was unstimulated (light orange filled histogram) or treated with 100 ng/mL Recombinant Human IL‑4 (Catalog # 204-IL) for 20 minutes, then stained with Rabbit Anti-Human Phospho‑STAT6 (Y641) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3717, dark orange filled histogram) or isotype control antibody (Catalog # AB‑105‑C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
Detection of Human Phospho-STAT6 (Y641) by Western Blot.
Western blot shows lysates of U937 human histiocytic lymphoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IL‑4 (Catalog # 204-IL) for 30 minutes. Gels were loaded with 20 μg of cytoplasmic (Cyto) and 10 μg of nuclear extracts (Nuc). PVDF membrane was probed with 0.5 µg/mL Rabbit Anti-Human Phospho-STAT6 (Y641) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3717) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-STAT6 (Y641) was detected at approximately 110 kDa (as indicated). For additional reference, the membrane was stripped and reprobed with 0.5 µg/mL Human STAT6 Monoclonal Antibody (lower panel, Catalog # MAB2167). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-STAT6 (Y641) by Simple WesternTM.
Simple Western lane view shows lysates of U937 human histiocytic lymphoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IL‑4 (Catalog # 204-IL) for 30 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-STAT6 (Y641) at approximately 101 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-STAT6 (Y641) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3717). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Preparation and Storage
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C, as supplied.
1 month, 2 to 8 °C under sterile conditions after opening.
6 months, -20 to -70 °C under sterile conditions after opening.
Signal Transducer and Activator of Transcription 6 (STAT6) mediates the signaling of cytokines such as IL-4 and IL-13. STAT6 acts as a signal transducer in the cytoplasm and, upon phosphorylation at Y641, translocates to the nucleus and binds to the DNA consensus site TTCN4GAA. Knockout studies in mice suggest that STAT6 functions in differentiation of T helper 2 (Th2) cells, expression of cell surface markers, and class switch of immunoglobulins.
Signal Transducer and Activator of Transcription 6
Entrez Gene IDs:
6778 (Human); 20852 (Mouse); 362896 (Rat)
D12S1644; EC 188.8.131.52; EC 184.108.40.206; IL-4 Stat; IL-4-STAT; signal transducer and activator of transcription 6; signal transducer and activator of transcription 6, interleukin-4 induced; STAT, interleukin4-induced; STAT6; STAT6B; STAT6C; transcription factor IL-4 STAT
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