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Signal Transduction Signaling Pathway Nuclear Signaling STATs
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-STAT3 antibody [EPR361] (ab109085)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (10)

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Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [EPR361] (ab109085)
  • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] (ab109085)
  • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
  • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
  • Flow Cytometry - Anti-STAT3 antibody [EPR361] (ab109085)
  • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
  • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] (ab109085)
  • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
  • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
  • Anti-STAT3 antibody [EPR361] (ab109085)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR361] to STAT3
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-STAT3 antibody [EPR361]
    See all STAT3 primary antibodies
  • Description

    Rabbit monoclonal [EPR361] to STAT3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1, HEK293, A431, Raji, HeLa, HaCaT, NIH/3T3, C2C12, and SH-SY5Y cell lysates. Human, mouse, and rat brain, mouse heart, and rat kidney tissue lysates. ICC/IF: HeLa and A459 cells. IHC-P: Human pancreas tissue. Flow Cyt: HeLa cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR361
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Stem Cells
    • Embryonic Stem Cells
    • Intracellular
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • STATs
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • STAT family
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Transcription factors
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Surface molecules

Associated products

  • Alternative Versions

    • Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    • Alexa Fluor® 488 Anti-STAT3 antibody [EPR361] (ab223404)
    • Alexa Fluor® 647 Anti-STAT3 antibody [EPR361] (ab223405)
  • Assay kits

    • STAT3 Transcription Factor Assay Kit (Colorimetric) (ab207229)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human STAT3 knockout HeLa cell line (ab255436)
  • KO cell lysates

    • Human STAT3 knockout HeLa cell lysate (ab263797)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • Raji whole cell lysate (ab30124)
  • Recombinant Protein

    • Recombinant Human STAT3 protein (ab43618)

Applications

The Abpromise guarantee

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 IHC antigen retrieval protocol.

ICC/IF
1/150.
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 IHC antigen retrieval protocol.

ICC/IF
1/150.
  • Application notes
    Is unsuitable for IP.
  • 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.
    • Target information above from: UniProt accession P40763 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 6774 Human
      • Entrez Gene: 20848 Mouse
      • Entrez Gene: 25125 Rat
      • Omim: 102582 Human
      • SwissProt: P40763 Human
      • SwissProt: P42227 Mouse
      • SwissProt: P52631 Rat
      • Unigene: 463059 Human
      • Unigene: 249934 Mouse
      • Unigene: 473190 Mouse
      • Unigene: 10247 Rat
      see all
    • 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

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

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : STAT3 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 88 kDa
      Observed band size: 92 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab109085 observed at 92 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab109085 was shown to react with STAT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255436 (knockout cell lysate ab263797) was used. Wild-type HeLa and STAT3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109085 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [EPR361] (ab109085)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [EPR361] (ab109085)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] (ab109085)
      Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] (ab109085)

      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.

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

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : HaCaT (Human skin keratinocyte) whole cell lysate
      Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
      Lane 4 : C2C12 (Mouse myoblasts cell line) whole cell lysate
      Lane 5 : Human brain lysate
      Lane 6 : Mouse brain lysate
      Lane 7 : Rat brain lysate
      Lane 8 : Rat liver lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 88 kDa
      Observed band size: 92 kDa why is the actual band size different from the predicted?
      Additional bands at: 32 kDa, 40 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 50 seconds


      Blocking/Diluting buffer: 5% NFDM/TBST.

      Rabbit monoclonal [EPR16891] to GAPDH (ab181602) used as loading control.

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

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : STAT3 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : HEK293 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 88 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab109085 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Ab109085 was shown to specifically react with STAT3 in wild-type cells as signal was lost in STAT3 knockout HAP1 cells. 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.

    • Flow Cytometry - Anti-STAT3 antibody [EPR361] (ab109085)
      Flow Cytometry - Anti-STAT3 antibody [EPR361] (ab109085)

      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.

    • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
      Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
      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
      All lanes : 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

    • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] (ab109085)
      Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] (ab109085)

      Immunofluorescence staining of A549 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.

    • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
      Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
      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 (Human neuroblastoma cell line from bone marrow) cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : 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

    • Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
      Western blot - Anti-STAT3 antibody [EPR361] (ab109085)
      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

    • Anti-STAT3 antibody [EPR361] (ab109085)
      Anti-STAT3 antibody [EPR361] (ab109085)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (10)

    Publishing research using ab109085? Please let us know so that we can cite the reference in this datasheet.

    ab109085 has been referenced in 10 publications.

    • Wang N  et al. Yeast ß-D-glucan exerts antitumour activity in liver cancer through impairing autophagy and lysosomal function, promoting reactive oxygen species production and apoptosis. Redox Biol 32:101495 (2020). PubMed: 32171725
    • Li Y  et al. The anthelmintic flubendazole blocks human melanoma growth and metastasis and suppresses programmed cell death protein-1 and myeloid-derived suppressor cell accumulation. Cancer Lett 459:268-276 (2019). PubMed: 31128215
    • He J  et al. Interleukin-9 promotes tumorigenesis through augmenting angiogenesis in non-small cell lung cancer. Int Immunopharmacol 75:105766 (2019). PubMed: 31352324
    • Silva C  et al. Selective pro-apoptotic and antimigratory effects of polyphenol complex catechin:lysine 1:2 in breast, pancreatic and colorectal cancer cell lines. Eur J Pharmacol 859:172533 (2019). PubMed: 31301308
    • Tan C  et al. Extensive protein S-nitrosylation associated with human pancreatic ductal adenocarcinoma pathogenesis. Cell Death Dis 10:914 (2019). PubMed: 31801946
    • Cao P  et al. MiR-15b is a key regulator of proliferation and apoptosis of chondrocytes from patients with condylar hyperplasia by targeting IGF1, IGF1R and BCL2. Osteoarthritis Cartilage N/A:N/A (2018). PubMed: 30521861
    • Ohkuro M  et al. Calreticulin and integrin alpha dissociation induces anti-inflammatory programming in animal models of inflammatory bowel disease. Nat Commun 9:1982 (2018). PubMed: 29773794
    • Cui X  et al. Baicalein represses TGF-ß1-induced fibroblast differentiation through the inhibition of miR-21. Toxicol Appl Pharmacol 358:35-42 (2018). PubMed: 30201452
    • Cheng ZX  et al. MicroRNA-132 induces temozolomide resistance and promotes the formation of cancer stem cell phenotypes by targeting tumor suppressor candidate 3 in glioblastoma. Int J Mol Med 40:1307-1314 (2017). PubMed: 28901390
    • 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 . PubMed: 27567678

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-STAT3 antibody [EPR361]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 17 2014

    Immunohistochemistry (Frozen sections) abreview for Anti-STAT3 antibody [EPR361]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 28 2014

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