Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

Overview

  • Product nameAnti-STAT5 (phospho Y694) antibody [E208]
    See all STAT5 primary antibodies
  • Description
    Rabbit monoclonal [E208] to STAT5 (phospho Y694)
  • SpecificityThe antibody only detects Stat5 phosphorylated on Tyrosine 694.
  • Tested applicationsSuitable for: WB, IHC-Pmore details
    Unsuitable for: ICC/IF or IP
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human STAT5 (phospho Y694).

  • Positive control
    • WB: A431 cell lysate treated with epidermal growth factor. IHC-P: Human breast carcinoma tissue.
  • 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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab32364 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 90 kDa (predicted molecular weight: 90 kDa).
IHC-P 1/50.
  • Application notesIs unsuitable for ICC/IF or IP.
  • Target

    • FunctionCarries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription.
    • Sequence similaritiesBelongs to the transcription factor STAT family.
      Contains 1 SH2 domain.
    • Post-translational
      modifications
      Tyrosine phosphorylated in response to IL-2, IL-3, IL-7, IL-15, GM-CSF, growth hormone, prolactin, erythropoietin and thrombopoietin. Tyrosine phosphorylation is required for DNA-binding activity and dimerization. Serine phosphorylation is also required for maximal transcriptional activity.
    • Cellular localizationCytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation.
    • Information by UniProt
    • Database links
    • Alternative names
      • MGF antibody
      • Signal transducer and activator of transcription 5A antibody
      • Signal Transducer and Activator of Transcription 5B antibody
      • STA5A_HUMAN antibody
      • STAT 5A antibody
      • STAT 5B antibody
      • STAT5 antibody
      • STAT5A antibody
      • STAT5B antibody
      • Transcription factor STAT5A antibody
      • Transcription factor STAT5B antibody
      see all

    Anti-STAT5 (phospho Y694) antibody [E208] images

    • Serially diluted ab32364 was bound to immobilised phospho peptide (P) - or control peptides ((C), 1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 1: 50,000) and signal was developed with TMB substrate

    • All lanes : Anti-STAT5 (phospho Y694) antibody [E208] (ab32364) at 1/1000 dilution (purified)

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

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size : 90 kDa
      Observed band size : 90 kDa

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-STAT5 (phospho Y694) antibody [E208] (ab32364) at 1/1000 dilution (unpurified)

      Lane 1 : A431 cell lysate, untreated
      Lane 2 : A431 cell lysate, treated with EGF
      Lane 3 : A431 cell lysate, treated with phosphatase


      Predicted band size : 90 kDa
      Observed band size : 90 kDa
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human infiltrating duct carcinoma of breast tissue labelling STAT5 (phospho Y694) with purified ab32364 at 1/50. 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma using unpurified ab32364 at a 1/50 dilution.

    References for Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

    This product has been referenced in:
    • Yang TY  et al. A multiple reaction monitoring (MRM) method to detect Bcr-Abl kinase activity in CML using a peptide biosensor. PLoS One 8:e56627 (2013). WB ; Human . Read more (PubMed: 23437189) »
    • Caldarelli A  et al. A genome-wide RNAi screen identifies proteins modulating aberrant FLT3-ITD signaling. Leukemia 27:2301-10 (2013). WB ; Human . Read more (PubMed: 23508117) »

    See all 8 Publications for this product

    Product Wall

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Rat Tissue sections (Mesenteric LN)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA pH 9.0
    Permeabilization No
    Specification Mesenteric LN
    Blocking step Casein in PBS as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Nov 30 2016

    I can confirm that the sequence around the phosphorylation site for STAT5A phospho Y694 and STAT5B phospho Y699 is 100% homolog:


    5A: VDGYPVKP

    5B: VDGYPVKP


    Therefore any antibody that binds STAT5 (phospho Y694) will ...

    Read More
    Application Immunocytochemistry/ Immunofluorescence
    Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
    Sample Human Cell (Hela cells)
    Specification Hela cells
    Permeabilization Yes - citrate buffer, pH 6.0
    Fixative Methanol
    Username

    Abcam user community

    Verified customer

    Submitted Mar 23 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6,0
    Sample Human Tissue sections (skin)
    Specification skin
    Permeabilization Yes - 0,1% Saponin/PBS
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Mar 17 2015

    Application Western blot
    Loading amount 15 µg
    Gel Running Conditions Reduced Non-Denaturing (Native) (5-16% gel)
    Sample Human Cell lysate - whole cell (Jurkat and Hela cells)
    Specification Jurkat and Hela cells
    Treatment see below
    Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Mar 17 2015

    Thank you for contacting us. For a positive control for human cells, I recommend the nuclear lysate of EGF-treated A431 cells which we have in our catalogue as ab14635: Click here (or use the following: http://www.abcam.com/index.html?datasheet=1...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (Lung cancer cell lines)
    Loading amount 100000 cells
    Specification Lung cancer cell lines
    Treatment 0, 0.25, 0.5, 1, 2, 4 uM dasatinib for 3 hours
    Gel Running Conditions Reduced Denaturing (4-12% NuPage gels)
    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 Apr 12 2011

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (Colon cancer)
    Loading amount 40 µg
    Specification Colon cancer
    Gel Running Conditions Non-reduced Denaturing
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 21 2010

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