Validated using a knockout cell line
Recombinant
RabMAb

Anti-STAT3 antibody [EPR361] (ab109085)

Overview

  • Product nameAnti-STAT3 antibody [EPR361]
    See all STAT3 primary antibodies
  • Description
    Rabbit monoclonal [EPR361] to STAT3
  • Tested applicationsSuitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human STAT3 aa 650-750.

  • Positive control
    • WB: HAP1, HEK293, A431, Raji, HeLa, and SH-SY5Y cell lysates and mouse heart and rat brain tissue lysates. ICC/IF: HeLa and A459 cells. IHC-P: Human pancreas tissue. Flow Cyt: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    A trial size is available to purchase for this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109085 in the following tested applications.

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

Application Abreviews Notes
Flow Cyt 1/300.
WB 1/1000 - 1/10000. Predicted molecular weight: 88 kDa.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/150.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionSignal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1 (PubMed:17344214). Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation (PubMed:18242580). Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity.
    • Tissue specificityHeart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
    • Involvement in diseaseHyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
      Autoimmune disease, multisystem, infantile-onset
    • Sequence similaritiesBelongs to the transcription factor STAT family.
      Contains 1 SH2 domain.
    • Post-translational
      modifications
      Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. 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. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling.
    • Cellular localizationCytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. 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. Identified in a complex with LYN and PAG1.
    • Information by UniProt
    • Database links
    • Alternative names
      • 1110034C02Rik antibody
      • Acute Phase Response Factor antibody
      • Acute-phase response factor antibody
      • ADMIO antibody
      • APRF antibody
      • AW109958 antibody
      • DNA binding protein APRF antibody
      • FLJ20882 antibody
      • HIES antibody
      • MGC16063 antibody
      • Signal transducer and activator of transcription 3 (acute phase response factor) antibody
      • Signal transducer and activator of transcription 3 antibody
      • STAT 3 antibody
      • Stat3 antibody
      • STAT3_HUMAN antibody
      see all

    Anti-STAT3 antibody [EPR361] images



    • Predicted band size : 88 kDa

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: STAT3 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HEK293 whole cell lysate (20 µg)

       

      Lanes 1 - 4: Merged signal (red and green). Green - ab109085 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

       

      ab109085 was shown to specifically react with STAT3 when STAT3 knockout samples were used. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Ab109085 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling STAT3 with Purified ab109085 at 1/250 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    • Immunohistochemical staining of paraffin embedded human pancreas with purified ab109085 at a working dilution of 1 in 100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling STAT3 (red) with ab109085 at a 1/300 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

    • All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/2000 dilution (purified)

      Lane 1 : Mouse heart tissue lysate
      Lane 2 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 88 kDa
      Observed band size : 88 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunofluorescence staining of A459 cells with purified ab109085 at a working dilution of 1 in 150, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit (ab150078), used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab109085 was used at a dilution of 1/200 followed by an Alexa Fluor® 488 goat anti-mouse antibody at a dilution of 1/500.

    • All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/2000 dilution (purified)

      Lane 1 : A431 cell lysate
      Lane 2 : Raji cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : SH-S5SY cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1000 mg/ml

      Predicted band size : 88 kDa
      Observed band size : 88 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/1000 dilution (unpurified)

      Lane 1 : A431 cell lysate
      Lane 2 : Raji cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : SH-SY5Y cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 88 kDa

    References for Anti-STAT3 antibody [EPR361] (ab109085)

    This product has been referenced in:
    • Mao S  et al. miR-17-92 facilitates neuronal differentiation of transplanted neural stem/precursor cells under neuroinflammatory conditions. J Neuroinflammation 13:208 (2016). WB ; Mouse . Read more (PubMed: 27567678) »

    See 1 Publication for this product

    Product Wall

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (4-15% TRIS-Glycine SDS PAGE)
    Sample Mouse Tissue lysate - whole (1) mouse embryonic spinal cord E11, 2) HEK293H)
    Specification 1) mouse embryonic spinal cord E11, 2) HEK293H
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

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    Verified customer

    Submitted Nov 17 2014

    Application Immunohistochemistry (Frozen sections)
    Blocking step 0.25%TritonX100, 0.2%Gelatin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 25°C
    Sample Mouse Tissue sections (mouse embryonic spinal cord E11)
    Specification mouse embryonic spinal cord E11
    Permeabilization Yes - 0.25%TritonX100
    Fixative Paraformaldehyde
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    Verified customer

    Submitted Jul 28 2014

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