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RabMAb

Anti-STAT3抗体[EPR787Y] (ab68153)

概述

  • 产品名称
    Anti-STAT3抗体[EPR787Y]
    参阅全部 STAT3 一抗
  • 描述
    兔单克隆抗体[EPR787Y] to STAT3
  • 经测试应用
    适用于: ICC/IF, WB, Flow Cyt, IHC-Pmore details
    不适用于: IP
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human STAT3 aa 1-100 (N terminal).
    Database link: P40763

  • 阳性对照
    • WB: Rat and mouse heart tissue lysates, HAP1, A431 and Raji cell lysates. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: Raji cells.
  • 常规说明

     

     

    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.

性能

应用

Our Abpromise guarantee covers the use of ab68153 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF 1/200.

For unpurified use at 1/140.

WB 1/1000 - 1/2000. Detects a band of approximately 75, 88 kDa (predicted molecular weight: 88 kDa).
Flow Cyt 1/30 - 1/50.
IHC-P 1/200.

For unpurified use at 1/140.

  • 应用说明
    Is unsuitable for IP.
  • 靶标

    • 功能
      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.
    • 组织特异性
      Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
    • 疾病相关
      Hyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
      Autoimmune disease, multisystem, infantile-onset
    • 序列相似性
      Belongs to the transcription factor STAT family.
      Contains 1 SH2 domain.
    • 翻译后修饰
      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.
    • 细胞定位
      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
    • 数据库链接
    • 别名
      • 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

    图片



    • 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 - ab68153 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.

       

      Ab68153 detected the expected band for STAT3 in wild-type cells along with additional cross-reactive bands. The band was not seen in STAT3 knockout HAP1 cells. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Ab68153 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 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.

    • All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/2000 dilution (unpurified)

      Lane 1 : Rat heart tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : A431 cell lysate

      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 : 92 kDa (why is the actual band size different from the predicted?)

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue sections labelling STAT3 with purified ab68153 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling STAT3 (green) with purified ab68153 at 1/200. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    • ab68153 staining STAT3 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/30. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

      Isoytype control: Rabbit monoclonal IgG (Black)

      Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    • Flow cytometry analysis of Raji cells labelling STAT3 with unpurified ab68153 at 1/30 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. A rabbit monoclonal IgG was used as the isotype control (green).

    • All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/2000 dilution (purified)

      Lane 1 : Rat heart tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : A431 cell lysate

      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 : 92 kDa (why is the actual band size different from the predicted?)

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/500 dilution (unpurified)

      Lane 1 : A431 cell lysate
      Lane 2 : Raji cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 88 kDa
      Observed band size : 92 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 75 kDa. We are unsure as to the identity of these extra bands.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue sections labelling STAT3 with unpurified ab68153 at 1/140. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling STAT3 (green) with unpurified ab68153 at 1/140. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    • Flow cytometry analysis of Raji cells labelling STAT3 with purified ab68153 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. A rabbit monoclonal IgG was used as the isotype control (green).

    文献

    This product has been referenced in:
    • Li R  et al. STAT3 and NF-?B are Simultaneously Suppressed in Dendritic Cells in Lung Cancer. Sci Rep 7:45395 (2017). Read more (PubMed: 28350008) »
    • Wu Z  et al. Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells. Sci Rep 7:2210 (2017). Read more (PubMed: 28526849) »

    See all 14 Publications for this product

    客户评价及客户问答

    Filter by Application

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    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (CT26-WT tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9
    Permeabilization
    No
    Specification
    CT26-WT tumor
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Jun 07 2016

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing (12.5%)
    Loading amount
    1e+006 cells
    Specification
    HeLa
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Abcam user community

    Verified customer

    提交于 Jan 12 2016

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Colon)
    Gel Running Conditions
    Reduced Denaturing (12.5%)
    Loading amount
    1e+006 cells
    Specification
    Colon
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Abcam user community

    Verified customer

    提交于 Jan 12 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

    提交于 Aug 27 2015

    Application
    Western blot
    Loading amount
    20 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
    Username

    Abcam user community

    Verified customer

    提交于 Apr 06 2015

    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
    Username

    Abcam user community

    Verified customer

    提交于 Aug 01 2014

    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
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Verified customer

    提交于 Jul 28 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
    Username

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    提交于 Jun 09 2014

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

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