Detection of Phospho-STAT1 in IFN-alpha -treated Daudi Human Cell Line by Flow Cytometry.
Daudi human Burkitt's lymphoma cell line was unstimulated (light orange filled histogram) or treated with 500 units/mL Recombinant Human IFN‑ alpha (Catalog # 11100-1) for 20 minutes (dark orange filled histogram), then stained with Rabbit Anti-Human/Mouse Phospho‑STAT1 (Y701) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF2894) or 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-STAT1 (Y701) by Western Blot.
Western blot shows lysates of Daudi human Burkitt's lymphoma cell line untreated (-) or treated (+) with Recombinant Human IFN-alpha 2a for 20 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse Phospho-STAT1 (Y701) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2894), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-STAT1 (Y701) at approximately 94 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Human Phospho-STAT1 (Y701) by Simple WesternTM.
Simple Western lane view shows lysates of Daudi human Burkitt's lymphoma cell line untreated (-) or treated (+) with Recombinant Human IFN‑ alpha A (Catalog # 11100-1) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-STAT1 (Y701) at approximately 86 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse Phospho-STAT1 (Y701) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2894). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. 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 reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
STAT1 (signal transducer and activator of transcription #1) is an 88 kDa member of the STAT family of cytoplasmic transcription factors. STAT members generally mediate cytokine, growth factor and hormone receptor signal transduction. STAT1 is associated with type I and II interferon signaling. All STATs contain an N-terminal oligomerization domain, a DNA-binding domain, and an SH2-association region. STAT1 is phosphorylated at Y701 by receptor-associated Janus kinases (JAKs) leading to STAT1 dimerization and subsequent translocation to the nucleus to activate gene transcription.
Signal Transducer and Activator of Transcription 1
Entrez Gene IDs:
6772 (Human); 20846 (Mouse)
CANDF7; DKFZp686B04100; ISGF-3; ISGF-3,91kD; signal transducer and activator of transcription 1, 91kDa; signal transducer and activator of transcription-1; STAT1; STAT91
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