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    stat3-antibody-e121-21-ab32500.pdf

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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 [E121-21] (ab32500)

  • Datasheet
  • SDS
Reviews (3) Submit a question References (22)

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Western blot - Anti-STAT3 antibody [E121-21] (ab32500)
  • Flow Cytometry (Intracellular) - Anti-STAT3 antibody [E121-21] (ab32500)
  • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [E121-21] (ab32500)
  • Western blot - Anti-STAT3 antibody [E121-21] (ab32500)
  • Immunoprecipitation - Anti-STAT3 antibody [E121-21] (ab32500)
  • Anti-STAT3 antibody [E121-21] (ab32500)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E121-21] to STAT3
  • Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Primary
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Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (ab171358)
Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
Product image
Human STAT3 knockout HeLa cell line (ab255436)

View more associated products

Overview

  • Product name

    Anti-STAT3 antibody [E121-21]
    See all STAT3 primary antibodies
  • Description

    Rabbit monoclonal [E121-21] to STAT3
  • Host species

    Rabbit
  • Specificity

    The antibody only detects Stat3 without phosphorylation on Serine 727. It does not detect S727-phosphorylated Stat3.
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Horse, Cow
  • Immunogen

    Synthetic peptide corresponding to Human STAT3. A synthetic peptide corresponding to residues surrounding Ser727 of human Stat3.

  • Positive control

    • HAP1, A431 cell lysate ICC/IF: HeLa cells Flow Cyt (intra): HeLa cells. IP: 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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E121-21
  • 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
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • 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 [E121-21] - BSA and Azide free (ab171361)
    • Alexa Fluor® 488 Anti-STAT3 antibody [E121-21] (ab214632)
    • Alexa Fluor® 647 Anti-STAT3 antibody [E121-21] (ab214939)
  • Assay kits

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

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human STAT3 protein (ab43618)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32500 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
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 88 kDa).
ICC/IF (2)
1/50.

For unpurified use at 1/250 dilution

IP
1/20.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/100 dilution

Notes
WB
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 88 kDa).
ICC/IF
1/50.

For unpurified use at 1/250 dilution

IP
1/20.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/100 dilution

Application notes
Is unsuitable for IHC-P.

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: 508541 Cow
    • Entrez Gene: 6774 Human
    • Omim: 102582 Human
    • SwissProt: P61635 Cow
    • SwissProt: P40763 Human
    • Unigene: 463059 Human
    • 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 [E121-21] (ab32500)
      Western blot - Anti-STAT3 antibody [E121-21] (ab32500)
      All lanes : Anti-STAT3 antibody [E121-21] (ab32500) at 1/1000 dilution (Purified)

      Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
      Lane 2 : HaCaT (Human skin keratinocyte) whole cell lysate

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

      Predicted band size: 88 kDa

    • Flow Cytometry (Intracellular) - Anti-STAT3 antibody [E121-21] (ab32500)
      Flow Cytometry (Intracellular) - Anti-STAT3 antibody [E121-21] (ab32500)

      Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labelling STAT3 with Purified ab32500 at 1:20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [E121-21] (ab32500)
      Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [E121-21] (ab32500)

      Immunocytochemistry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling STAT3 with Purified ab32500 at 1:50 dilution (2.1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Western blot - Anti-STAT3 antibody [E121-21] (ab32500)
      Western blot - Anti-STAT3 antibody [E121-21] (ab32500)

      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 - ab32500 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.

       

      Ab32500 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. Ab32500 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.

    • Immunoprecipitation - Anti-STAT3 antibody [E121-21] (ab32500)
      Immunoprecipitation - Anti-STAT3 antibody [E121-21] (ab32500)

      ab32500 at 1/20 dilution (0.5 µg) immunoprecipitating STAT3 in HeLa whole cell lysate.

      Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab32500 & HeLa whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32500 in HeLa whole cell lysate

      For western blotting, ab32500 at 1/1000 and VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

      Blocking and diluting buffer: 5% NFDM /TBST.

    • Anti-STAT3 antibody [E121-21] (ab32500)
      Anti-STAT3 antibody [E121-21] (ab32500)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (22)

    Publishing research using ab32500? Please let us know so that we can cite the reference in this datasheet.

    ab32500 has been referenced in 22 publications.

    • Li R  et al. miR-21a inhibits decidual cell apoptosis by targeting Pdcd4. Genes Dis 8:171-180 (2021). PubMed: 33997164
    • Wang C  et al. Blocking the autocrine regulatory loop of Gankyrin/STAT3/CCL24/CCR3 impairs the progression and pazopanib resistance of clear cell renal cell carcinoma. Cell Death Dis 11:117 (2020). PubMed: 32051393
    • Pu Z  et al. Circular RNA circCUL3 Accelerates the Warburg Effect Progression of Gastric Cancer through Regulating the STAT3/HK2 Axis. Mol Ther Nucleic Acids 22:310-318 (2020). PubMed: 33230436
    • Liu Z  et al. STAT3-induced upregulation of long noncoding RNA HNF1A-AS1 promotes the progression of oral squamous cell carcinoma via activating Notch signaling pathway. Cancer Biol Ther 20:444-453 (2019). PubMed: 30404566
    • Yang L  et al. Interleukin-35 modulates the balance between viral specific CD4+CD25+CD127dim/- regulatory T cells and T helper 17 cells in chronic hepatitis B virus infection. Virol J 16:48 (2019). PubMed: 30992023
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-STAT3 antibody [E121-21] - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (brain)
    Permeabilization
    Yes - 0.05% Triton X-100/PBS
    Specification
    brain
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 24 2009

    Immunocytochemistry/ Immunofluorescence abreview for Anti-STAT3 antibody [E121-21] - ChIP Grade

    Poor
    Abreviews
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK cells)
    Specification
    HEK cells
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 5%
    Fixative
    Methanol
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Randal Moldrich

    Verified customer

    Submitted Jun 19 2006

    Immunocytochemistry/ Immunofluorescence abreview for Anti-STAT3 antibody [E121-21] - ChIP Grade

    Poor
    Abreviews
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Glia cell culture)
    Specification
    Glia cell culture
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 5%
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Randal Moldrich

    Verified customer

    Submitted Jun 19 2006

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