Recombinant
RabMAb

Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Overview

  • Product name
    Anti-STAT3 (phospho S727) antibody [E121-31]
    See all STAT3 primary antibodies
  • Description
    Rabbit monoclonal [E121-31] to STAT3 (phospho S727)
  • Specificity
    This antibody detects Stat3 phosphorylated on Serine 727. It does not crossreact with the phospho serines of other Stat proteins.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human, Macaque monkey
    Predicted to work with: Horse, Cow
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human STAT3 aa 700 to the C-terminus.
    Database link: P40763

  • Positive control
    • WB: A431 cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab32143 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
ICC/IF 1/500.

For unpurified, use 1/100.

WB 1/1000 - 1/10000. Detects a band of approximately 98 kDa (predicted molecular weight: 88 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

For unpurified, use 1/50.

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

Anti-STAT3 (phospho S727) antibody [E121-31] images

  • Purified ab32143 staining STAT3 (phospho S727) in A431 cells by Immunocytochemistry/ Immunofluorescence. 4% PFA-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labelled with ab32143 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing nuclear staining on A431 cell line. The red staining is ab7291 anti-Tubulin (mouse mAb), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody.

  • Immunohistochemical staining of paraffin embedded human astrocytoma with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. 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.
  • All lanes : Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143) at 1/5000 dilution (purified)

    Lane 1 : C6 treated with epidermal growth factor
    Lane 2 : untreated C6 whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size : 88 kDa
    Observed band size : 98 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • All lanes : purified at 5000

    Lane 1 : A431 treated with epidermal growth factor
    Lane 2 : untreated A431 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size : 88 kDa
    Observed band size : 98 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded rat cerebral cortex with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. 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.
  • Immunohistochemical staining of paraffin embedded mouse liver with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. 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.
  • Immunocytochemical/Immunofluorescence analysis of untreated, EGF treated and EGF + LP treated A431 cells labelling STAT3 (phospho S727) with ab32143 (left) and STAT3 with ab32500 (right) both at a dilution of 1/500.

    Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) (1/1000) was used as the secondary antibody (green). DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (1/200) was used as a counterstain (red).

    The green staining was increased and translocated from the cytoplasm into the nucleus in the EGF (ab9697 100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signal was decreased. For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment.

  • Dot Blot analysis of Lane 1: STAT3 (pS727) phospho peptide and Lane 2: STAT3 non-phospho peptide labeling STAT3 (phospho S727) with ab32143 at 1/1000 dilution (0.009 μg/ml). 5% NFDM /TBST was used as the diluting and blocking buffer and concentration. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100,000 dilution. Exposure time: 10 seconds.

  • All lanes : Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143) at 1/1000 dilution (unpurified)

    Lane 1 : A431 cell lysate
    Lane 2 : A431 + EGF cell lysate


    Predicted band size : 88 kDa
    Observed band size : 98 kDa (why is the actual band size different from the predicted?)
  • IHC-P analysis of brain astrocytoma using unpurified ab32143 at 1/50 dilution.

References for Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

This product has been referenced in:
  • Li J  et al. Correlative study on the JAK-STAT/PSMß3 signal transduction pathway in asthenozoospermia. Exp Ther Med 13:127-130 (2017). Read more (PubMed: 28123480) »
  • Murphy MJ  et al. Leukemia Inhibitory Factor is Necessary for Ovulation in Female Rhesus Macaques. Endocrinology N/A:en20161283 (2016). IHC ; Macaque monkey . Read more (PubMed: 27571132) »

See all 8 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (pancreatic fiborblasts)
Gel Running Conditions
Reduced Denaturing (10% PAGE)
Loading amount
20 µg
Specification
pancreatic fiborblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 01 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
No
Specification
mouse pancreas
Blocking step
casein as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 4°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 01 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Esophageal cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Tris - 1mM EDTA - 0.05% Tween-20, pH 9.0
Permeabilization
No
Specification
Esophageal cancer
Blocking step
(agent) for 20 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde
Username

Tatiana Karakasheva

Verified customer

Submitted Mar 04 2016

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

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