Recombinant
RabMAb

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

Overview

  • Product name
    Anti-STAT3 (phospho Y705) antibody [EP2147Y]
    See all STAT3 primary antibodies
  • Description
    Rabbit monoclonal [EP2147Y] to STAT3 (phospho Y705)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, Dot blot, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human STAT3 aa 650-750 (phospho Y705). The exact sequence is proprietary.
    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. IP: A431 cells treated with EGF.
  • General notes

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    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.

    This product is a recombinant rabbit monoclonal 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 1/2000 - 1/20000. 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 IHC antigen retrieval protocols.

Dot blot 1/1000.
ICC/IF 1/500.

Target

  • Function
    Signal 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 specificity
    Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
  • Involvement in disease
    Hyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
    Autoimmune disease, multisystem, infantile-onset
  • Sequence similarities
    Belongs 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 localization
    Cytoplasm. 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

Images

  • All lanes : Anti-STAT3 (phospho Y705) antibody [EP2147Y] (ab76315) at 1/20000 dilution (purified)

    Lane 1 : Un-treated HeLa (Human epithelial cell line from cervix adenocarcinoma )cell lysate
    Lane 2 : HeLa cell lysate - treated with IFN-a

    Lysates/proteins at 10 µg per lane.

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

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labeling 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.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) +/- IFN-α (50 ng/mL, 5 minutes) cells labelling STAT3 (phospho Y705) with ab76315 at 1/500 (4.3 μg/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1500) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.

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

  • 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 dilution buffer was 5% NFDM/TBST. The secondary antibody used was ab97051 peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/100,000.

  • Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue using  untreated (left) or alkaline phosphatase-treated (right) labeling STAT3 (phospho Y705) with ab76315 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H& L (HRP) (ab97051) at 1/500 dilution.

    Counter stained with Hematoxylin.

  • ab76315 (purified) at 1/30 immunoprecipitating STAT3 (phospho Y705) in A431 (Human epidermoid carcinoma cell line) 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/Dilution buffer: 5% NFDM/TBST.

     

References

This product has been referenced in:
  • Hanna M  et al. Association between physical activity and the expression of mediators of inflammation in normal breast tissue among premenopausal and postmenopausal women. Cytokine 102:151-160 (2018). Read more (PubMed: 29102166) »
  • Chen L  et al. Combination of gemcitabine and erlotinib inhibits recurrent pancreatic cancer growth in mice via the JAK-STAT pathway. Oncol Rep 39:1081-1089 (2018). Read more (PubMed: 29328487) »

See all 95 Publications for this product

Customer reviews and Q&As

Filter by Application

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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
Username

Abcam user community

Verified customer

Submitted Jul 07 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Feb 05 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (stomach tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
stomach tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Feb 05 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Feb 05 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA 70C 2hr
Permeabilization
No
Specification
Colon
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 23 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Colon
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 28 2017

Application
ELISA
Sample
Human Cell (HEK293 cells stably overexpressing the human Lepti)
Specification
HEK293 cells stably overexpressing the human Lepti
Blocking step
proprietary blocking buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Type
Other
Username

Abcam user community

Verified customer

Submitted Sep 12 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Lung)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
22 µg
Specification
Lung
Blocking step
Odyssey TBS Block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C
Username

Pete Serbedzija

Verified customer

Submitted Jul 28 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human esophageal cancer subcutaneous xenograft gro)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.0
Specification
human esophageal cancer subcutaneous xenograft gro
Blocking step
Non-Abcam blocking agent as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde
Username

Tatiana Karakasheva

Verified customer

Submitted Feb 22 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (human normal oesopheal cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
10 nM PMA
Specification
human normal oesopheal cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Aug 27 2015

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