Anti-STAT3 (phospho S727) antibody (ab86430)

Overview

  • Product nameAnti-STAT3 (phospho S727) antibodySee all STAT3 primary antibodies ...
  • Description
    Rabbit polyclonal to STAT3 (phospho S727)
  • Tested applicationsICC/IF, WB, IHC-FoFr more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 700 to the C-terminus of Human STAT3.

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: A431; HeLa; NIH3T3.

Properties

Applications

Our Abpromise guarantee covers the use of ab86430 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ICC/IF Use a concentration of 1 µg/ml.
WB 1/250. Detects a band of approximately 88 kDa (predicted molecular weight: 88 kDa).
IHC-FoFr 1/300.

Target

  • FunctionTranscription factor that binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA.
  • Tissue specificityHeart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
  • Involvement in diseaseDefects in STAT3 are the cause of hyperimmunoglobulin E recurrent infection syndrome autosomal dominant (AD-HIES) [MIM:147060]; also known as hyper-IgE syndrome or Job syndrome. AD-HIES is a rare disorder of immunity and connective tissue characterized by immunodeficiency, chronic eczema, recurrent Staphylococcal infections, increased serum IgE, eosinophilia, distinctive coarse facial appearance, abnormal dentition, hyperextensibility of the joints, and bone fractures.
  • Sequence similaritiesBelongs to the transcription factor STAT family.
    Contains 1 SH2 domain.
  • Post-translational
    modifications
    Tyrosine phosphorylated upon stimulation with EGF (By similarity). Tyrosine phosphorylated in response to IL-6, IL-11, CNTF, LIF, CSF-1, EGF, PDGF, IFN-alpha and OSM. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus.
  • Cellular localizationCytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3.
  • Target information above from: UniProt accession P40763 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • 1110034C02Rik antibody
    • Acute Phase Response Factor antibody
    • Acute-phase response factor antibody
    • APRF antibody
    • AW109958 antibody
    • DNA binding protein APRF antibody
    • FLJ20882 antibody
    • HIES antibody
    • MGC16063 antibody
    • Signal transducer and activator of transcription 3 antibody
    • STAT 3 antibody
    • Stat3 antibody
    • STAT3_HUMAN antibody
    see all

Anti-STAT3 (phospho S727) antibody images

  • All lanes : Anti-STAT3 (phospho S727) antibody (ab86430) at 1/250 dilution

    Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) EGF Stimulated Whole Cell Lysate
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunising Peptide at 1/250 dilution
    Lane 5 : A431 (Human epithelial carcinoma cell line) EGF Stimulated Whole Cell Lysate with Immunising Peptide at 1/250 dilution
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunising Peptide at 1/250 dilution
    Lane 7 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Non-Modified Control Peptide at 1/250 dilution
    Lane 8 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab52255) with Non-Modified Control Peptide at 1/250 dilution
    Lane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Non-Modified Control Peptide at 1/250 dilution

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 88 kDa
    Observed band size : 88 kDa
    Additional bands at : 50 kDa,60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
  • ICC/IF image of ab86430 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab86430 at 1µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in Hek293, HepG2, and MCF-7 formaldehyde (4%, 10min) fixed cells at 1ug/ml, and also in HeLa, Hek293, HepG2, and MCF-7 Methanol (100%, 5min) fixed cells at 1ug/ml.

  • IHC- FoFr image of STAT3 (phospho S727) staining using rat hypothalamus at low (left) and high (right) magnification. The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the spinal cord were cryoprotected in sucrose 30% overnight. Spinal cords were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

    See Abreview

  • Anti-STAT3 (phospho S727) antibody (ab86430) at 1 µg/ml + NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 88 kDa
    Observed band size : 88 kDa
    Additional bands at : 180 kDa,40 kDa,70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

References for Anti-STAT3 (phospho S727) antibody (ab86430)

ab86430 has not yet been referenced specifically in any publications.

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Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain - Hypothalamus)
Specification Brain - Hypothalamus
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Mar 20 2012

Thank you for contacting us.  Both ab86132 and ab86430 are raised in rabbit and bind to the phosphorylated serine 727 in STAT1 and STAT3 respectively.  I hope this helps, please let me know if you need any additional information.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"