Detection of STAT3-regulated Genes by Chromatin Immunoprecipitation.
Jurkat human acute T cell leukemia cell line treated with 1000 U/mL Recombinant Human IFN‑ alpha Universal Type I (Catalog # 11200-1) for 1 hour was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. STAT3/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse/Rat STAT3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1799) or control antibody (Catalog # AB‑108‑C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The c-myc promoter was detected by standard PCR.
Immunoprecipitation of Human STAT3.
STAT3 was immunoprecipitated from 500 μg of Daudi human Burkitt's lymphoma cell line cytoplasmic and nuclear lystates following incubation with 2 µg Goat Anti-Human/Mouse/Rat STAT3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1799) for overnight at 4 ºC. STAT3-antibody complexes were absorbed using Protein G agarose (Invitrogen). Immunoprecipitated STAT3 was detected by Western blot using 0.1 µg/mL Human STAT3 Monoclonal Antibody (Catalog # MAB1799). View our recommended buffer recipes for immunoprecipitation.
STAT3 in D3 Mouse Cell Line.
STAT3 was detected in immersion fixed D3 mouse embryonic stem cell line using Goat Anti-Human/Mouse/Rat STAT3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1799) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001). Tubulin was also detected using Tubulin Antibody (Catalog # NB600-506; Novus Biologicals) and stained using the NorthernLights™ 493-conjugated Anti-Rat IgG Secondary Antibody (green; Catalog # NL015). Cells were counterstained with DAPI (blue). Specific staining of STAT3 was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
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.
The STAT family of transcription factors, first described as key mediators of interferon signal transduction, are now recognized as signaling factors downstream of many cytokines, growth factors, and hormones. At least seven STATs exist: STAT1, 2, 3, 4, 5a, 5b, and 6. They are responsible for an array of cellular activities including regulating growth, survival, differentiation, motility, and the immune response. They are characterized structurally by the presence of the Src homology 2 (SH2) domain, and a C-terminal tyrosine phosphorylation site. Upon ligand binding, they are recruited to the activated receptor, via their SH2 domains, to receptor-associated tyrosine kinases of the Janus kinase family (JAK). Once phosphorylated by JAK, STATS are free to hetero- or homo-dimerize, and translocate to the nucleus where they bind DNA to regulate gene transcription.
Signal Transducer and Activator of Transcription 3
Entrez Gene IDs:
6774 (Human); 20848 (Mouse); 25125 (Rat)
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
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.
Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you
include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image.
Limited to verified customers in USA, Canada and Europe.
Read what people are saying who have used Human/Mouse/Rat STAT3 Antibody.
We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Immunoprecipitation.