Human/Mouse/Rat STAT3 Antibody MAB1799: R&D Systems

Human/Mouse/Rat STAT3 Antibody

(2 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat STAT3 in Western blots.
  • Source
    Monoclonal Mouse IgG2B Clone # 232209
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human STAT3
    Met1-Asn175
    Accession # P40763
  • Formulation
    Supplied as a 0.2 μm filtered solution in PBS. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunoprecipitation
    1-3 µg/500 µg cell lysate
    Daudi human Burkitt's lymphoma cell line, see our available Western blot detection antibodies
  • Immunocytochemistry
    3-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
  • Knockout Validated

    STAT3 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in STAT3 knockout HeLa cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Intracellular Staining by Flow Cytometry
Detection of STAT3 in Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cell line was stained with Mouse Anti-Human STAT3 Monoclonal Antibody (Catalog # MAB1799, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
Detection of Human STAT3 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and PC‑12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human STAT3 Monoclonal Antibody (Catalog # MAB1799) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for STAT3 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
STAT3 in HeLa Human Cell Line. STAT3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat STAT3 Monoclonal Antibody (Catalog # MAB1799) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Knockout Validated
STAT3 Specificity is Shown by Immunocytochemistry in KnockoutCell Line. STAT3 was detected in immersion fixed HeLa human cervicalepithelial carcinoma cell line treated with IFN-alpha 1, but is not detected inSTAT3 knockout (KO) HeLa cell line using Mouse Anti-Human/
Mouse/Rat STAT3Monoclonal Antibody (Catalog # MAB1799) at 1 µg/mL for 3 hours at roomtemperature. Cells were stained using the NorthernLights™ 557-conjugatedAnti-Mouse IgG Secondary Antibody (red; Catalog # NL007)and counterstained with DAPI (blue). Specific staining was localized tocytoplasm and nuclei. View our protocol for FluorescentICC Staining of Cells on Coverslips.
Knockout Validated
Western Blot Shows Human STAT3 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line, STAT1 knockout (KO) HeLa cell line, STAT2 KO HeLa cell line, STAT3 KO HeLa cell line, STAT5a KO HeLa cell line, STAT5b KO HeLa cell line, STAT6 KO HeLa cell line,. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat STAT3 Monoclonal Antibody (Catalog # MAB1799) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for STAT3 at approximately 80 kDa (as indicated) in the parental HeLacell line, but is not detectable in the STAT3 knockout HeLa cell line. GAPDH(Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Shipping
    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.
Background: STAT3
Signal Transducer and Activator of Transcription (STAT) proteins are transcription factors activated in response to cytokine, growth factor, or hormone receptor signaling. Janus kinases (JAKs) phosphorylate STAT proteins and induce dimerization. Homo- or heterodimers translocate to the nucleus where they bind to DNA and activate transcription.
  • Long Name:
    Signal Transducer and Activator of Transcription 3
  • Entrez Gene IDs:
    6774 (Human); 20848 (Mouse); 25125 (Rat)
  • Alternate Names:
    Acute-phase response factor; APRFMGC16063; DNA-binding protein APRF; FLJ20882; HIES; signal transducer and activator of transcription 3 (acute-phase responsefactor); signal transducer and activator of transcription 3; STAT3
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Proangiogenic tumor proteins as potential predictive or prognostic biomarkers for bevacizumab therapy in metastatic colorectal cancer.
    Authors: Bruhn M, Townsend A, Khoon Lee C, Shivasami A, Price T, Wrin J, Arentz G, Tebbutt N, Hocking C, Cunningham D, Hardingham J
    Int J Cancer, 2014;135(3):731-41.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: WB
  2. IL-6 cytoprotection in hyperoxic acute lung injury occurs via suppressor of cytokine signaling-1-induced apoptosis signal-regulating kinase-1 degradation.
    Authors: Kolliputi N, Waxman AB
    Am. J. Respir. Cell Mol. Biol., 2009;40(3):314-24.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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