Analysis Note

ControlSK-N-MC cellsmurine macrophage 4/4 cells, serum-starved for 16 hours then stimulated with Phorbol 12-myristate 13-acetate (PMA) for 10 minutes

Application

Use Anti-phospho-STAT3 (Tyr705) Antibody, clone 9E12 (Mouse Monoclonal Antibody) validated in ELISA, IP, WB to detect phospho-STAT3 (Tyr705) also known as Acute-phase response factor or DNA-binding protein APRF. This Stat3 Antibody has been validated in Human, Mouse and Canine samples.

Research Sub CategoryTranscription Factors

Research CategoryEpigenetics & Nuclear Function

Immunoprecipitation: An independent laboratory has reported 1-10 µg/mL of a previous lot of this antibody immunoprecipitated phospho-STAT3 (Tyr705) from a lysate of 106 vanadate-treated human A431 cells.

ELISA: 0.05 µg/mL of this antibody is recommended by an independent lab for ELISA testing. Test was conducted on a previous lot.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

STATs (signal transducers and activators of transcription) were originally discovered as two proteins (STAT1 and STAT2) which were involved in interferon-alpha (IFN-alpha) and IFN-gamma signal transduction. Since then, several additional STAT proteins have been identified (STAT3, 4, 5a, 5b, and 6). STATs undergo tyrosine phosphorylation in response to growth factor or cytokine signaling. This phosphorylation results in dimerization and translocation of STAT proteins to the nucleus. In some cases this process is mediated by JAK Kinases (Janus Kinases 1, 2, and 3). For maximum activation of these proteins, phosphorylation at specific tyrosine and serine residues may be required in STAT1 alpha, 3, 4, and 5. Specific functions of the various members of the STAT family are poorly understood. STAT3 has been shown to be activated by IFN-alpha but not IFN-beta. The transcription factors associated with STAT3 are c-Jun and cyclic AMP-responsive enhancer binding protein (CREB). Deletion of the STAT3 gene in knock-out mice was lethal at the early embryonic stage.

Immunogen

KLH-conjugated phosphopeptide containing the AP[pY]LK motif of STAT3

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-1059

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Thiophilic adsoption and size exclusion chromatography

Format: Purified

Quality

Routinely evaluated by immunoblot on lysates from human SK-N-MC cells stimulated with CNTF.

Western Blot Analysis:
0.1-1 µg/mL of this lot detected phospho-STAT3 (Tyr705) in lysates from human SK-NMC cells stimulated with CNTF.

Specificity

This antibody recognizes STAT3, Mr 92 kDa, phos-phorylated on tyrosine-705. Clone 9E12.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Target description

92 kDa