Description
Product Usage Information
For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits.
Western Blotting: 1:2000
Fluorescent Western: 1:2000
Simple Western™: 1:10 - 1:50
Immunoprecipitation: 1:100
IHC Leica Bond: 1:100 - 1:400
Immunohistochemistry (Paraffin): 1:100 - 1:400
Immunofluorescence (Immunocytochemistry): 1:100 - 1:200
Flow Cytometry (Fixed/Permeabilized): 1:100 - 1:400
Chromatin IP: 1:100
Chromatin IP-seq: 1:100
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier-free (BSA and azide free) version of this product see product # 73533 .
Protocol
Available protocols: Western Blotting, Fluorescent Western, Immunoprecipitation, IHC Leica Bond, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq
Specificity / Sensitivity
Phospho-Stat3 (Tyr705) (D3A7) Rabbit Monoclonal Antibody detects endogenous levels of Stat3 only when phosphorylated at tyrosine 705. This antibody does not cross-react with phospho-EGFR or the corresponding phospho-tyrosines of other Stat proteins.
Species Reactivity: Human, Mouse, Rat, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr705 of mouse Stat3.
Background
The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Research studies have shown that Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).
Alternate Names
Acute-phase response factor; ADMIO; ADMIO1; APRF; DNA-binding protein APRF; FLJ20882; HIES; MGC16063; Signal transducer and activator of transcription 3; signal transducer and activator of transcription 3 (acute-phase response factor); STAT3
