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.

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.

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')₂Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.

Detection of STAT3 in HeLa Human Cell Line by Flow Cytometry.

HeLa human cell line was stained with Mouse Anti-Human/Mouse/Rat STAT3 Monoclonal Antibody (Catalog # MAB1799, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram) followed by anti-Mouse IgG PE-conjugated secondary antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

STAT3 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

STAT3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with IFN-alpha 1, but is not detected in STAT3 knockout (KO) HeLa cell line using Mouse Anti-Human/ Mouse/Rat STAT3 Monoclonal Antibody (Catalog # MAB1799) at 1 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.

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 HeLa cell 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.

STAT3 Specificity is Shown by Flow Cytometry in Knockout Cell Line.

STAT3 knockout HeLa human cervical epithelial cell line was stained with Mouse Anti-Human/Mouse STAT3 Monoclonal Antibody (Catalog # MAB1799, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram) followed by anti-Mouse IgG PE-conjugated secondary antibody (Catalog # F0102B). No staining in the STAT3 knockout HeLa cell line was observed. To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

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Applications for Human/Mouse/Rat STAT3 Antibody

Application

Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells and HeLa human cervical epithelial carcinoma cell line

Immunoprecipitation

1-3 µg/500 µg cell lysate
Sample: Daudi human Burkitt's lymphoma cell line, see our available Western blot detection antibodies

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: HeLa or Jurkat human acute T cell leukemia cell line fixed with paraformaldehyde and permeabilized with methanol

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.

Western Blot

0.1 µg/mL
Sample: 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

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using MAB1799 in the following applications:

Immunocytochemistry/Immunofluorescence (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

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

Alternate Names

Acute-phase response factor, APRFMGC16063, DNA-binding protein APRF, FLJ20882, HIES, signal transducer and activator of transcription 3, signal transducer and activator of transcription 3 (acute-phase responsefactor)

Entrez Gene IDs

6774 (Human); 20848 (Mouse); 25125 (Rat)

Gene Symbol

STAT3

UniProt

P40763

Additional STAT3 Products

Product Documents for Human/Mouse/Rat STAT3 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human/Mouse/Rat STAT3 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human/Mouse/Rat STAT3 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB1799

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications