RabMAb

Anti-STAT3 (phospho Y705) antibody [EP2147Y] (ab76315)

Overview

  • Product nameAnti-STAT3 (phospho Y705) antibody [EP2147Y]
    See all STAT3 primary antibodies
  • Description
    Rabbit monoclonal [EP2147Y] to STAT3 (phospho Y705)
  • Tested applicationsWB, IP, IHC-P, ICC/IFmore details
  • 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 (phospho Y705).
    Database link: P40763
    (Peptide available as ab179551)

  • Positive control
    • WB: HeLa cell lysate treated with alpha-interferon. IHC-P: Human colon adenocarcinoma and kidney tissues. ICC/IF: HeLa cells. IP: A431 cells treated with EGF.
  • General notes

    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.

    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.

    A 40 µl trial size is available to purchase for this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76315 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
WB 1/200000. Predicted molecular weight: 88 kDa.Can be blocked with STAT3 (phospho Y705) peptide (ab179551).
IP 1/20.
IHC-P 1/50 - 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 1/100.

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.
  • 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 (acute phase response factor) antibody
    • Signal transducer and activator of transcription 3 antibody
    • Signal Transductor and Activator of Transcription 3 antibody
    • STAT 3 antibody
    • Stat3 antibody
    • STAT3_HUMAN antibody
    see all

Anti-STAT3 (phospho Y705) antibody [EP2147Y] images

  • Dot blot analysis of STAT3 single phospho peptide pY705 (lane 1) and STAT3 non-phospho peptide (lane 2) with ab76315 at 1/1000. Blocking and diluting buffer was 5% NFDM/TBST. The secondary antibody used was ab97051 Peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/100,000.

  • All lanes : Anti-STAT3 (phospho Y705) antibody [EP2147Y] (ab76315) at 1/200000 dilution (purified)

    Lane 1 : HeLa cell lysate - untreated
    Lane 2 : HeLa cell lysate - treated with IFN-a

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling STAT3 (phospho Y705) with purified ab76315 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue labelling STAT3 (phospho Y705) with unpurified ab76315 at 1/100.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling STAT3 (phospho Y705) with unpurified ab76315 at 1/100 dilution.

  • Unpurified ab76315 staining STAT3 (phospho Y705) in MDA-MB-231 cells treated with stattic (ab120952), by ICC/IF. Decrease in STAT3 (phospho Y705) expression correlates with increased concentration of stattic as described in literature.
    The cells were incubated at 37°C for 2h in media containing different concentrations of ab120952 (stattic) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with unpurified ab76315 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • ab76315 (purified) at 1/30 immunoprecipitating STAT3 (phospho Y705) in A431 cell lysate treated with EGF. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

References for Anti-STAT3 (phospho Y705) antibody [EP2147Y] (ab76315)

This product has been referenced in:
  • Wu J  et al. EGFR-STAT3 signaling promotes formation of malignant peripheral nerve sheath tumors. Oncogene 33:173-80 (2014). IHC ; Mouse . Read more (PubMed: 23318430) »
  • Turkington RC  et al. Fibroblast growth factor receptor 4 (FGFR4): a targetable regulator of drug resistance in colorectal cancer. Cell Death Dis 5:e1046 (2014). WB ; Human . Read more (PubMed: 24503538) »

See all 18 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (peripheral blood CD19+ B cells)
Loading amount 200 µg
Specification peripheral blood CD19+ B cells
Treatment (1) Untreated (2) 100 ng recombinant hIL-10 for 24hours
Gel Running Conditions Reduced Denaturing (10 % SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Verified customer

Submitted Jul 07 2010

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Spermophilus tridecemlineatus Tissue lysate - whole (brain)
Specification brain
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
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Submitted Oct 29 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (4–15%)
Sample Mouse Tissue lysate - whole (Lung)
Specification Lung
Blocking step BSA as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Jul 30 2014

Thank you for your enquiry.

I can confirm that for the western blot image on the datasheet for ab76315, the HeLa cellswere serum-starved overnight, and then treated with 1000 U/ml IFN alpha for 15min at 37 degrees C.

This may requ...

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Thank you for contacting us. Please be advised that the ab76315 is not a purified antibody; it is in tissue culture supernatant which may contains other proteins also. We generally do not determine the antibody concentration that is prepared as tissue ...

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