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    brachyury--bry-antibody-ab20680.pdf

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Stem Cells Lineage Markers Mesoderm
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Anti-Brachyury / Bry antibody (ab20680)

  • Datasheet
  • SDS
Reviews (5)Q&A (1)References (49)

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Immunocytochemistry/ Immunofluorescence - Anti-Brachyury / Bry antibody (ab20680)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody (ab20680)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody (ab20680)
  • Western blot - Anti-Brachyury / Bry antibody (ab20680)
  • Western blot - Anti-Brachyury / Bry antibody (ab20680)

Key features and details

  • Rabbit polyclonal to Brachyury / Bry
  • Suitable for: ICC/IF, IHC-P, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Brachyury / Bry antibody [EPR18113] (ab209665)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Brachyury / Bry antibody
    See all Brachyury / Bry primary antibodies
  • Description

    Rabbit polyclonal to Brachyury / Bry
  • Host species

    Rabbit
  • Specificity

    The immunogen used to raise this antibody has 81% homology with the Mouse Brachyury protein. Some customers have successfully used ab20680 on mouse samples, however we have not been successful detecting Brachyury in this species in our own testing and therefore cannot guarantee mouse reactivity. Please contact Abcam Scientific Support for more information.

  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: DogDoes not react with: Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human Brachyury/ Bry aa 250-350 (internal sequence) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab21992)

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Stem Cells
    • Lineage Markers
    • Mesoderm
    • Stem Cells
    • Hematopoietic Progenitors
    • Intracellular Molecules
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Developmental Biology
    • Lineage specification
    • Mesoderm

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • Human Brachyury / Bry peptide (ab21992)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Brachyury / Bry protein (ab114235)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab20680 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
Use at an assay dependent concentration.
IHC-P (3)
Use at an assay dependent concentration.
WB (2)
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
Notes
ICC/IF
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).

Target

  • Function

    Involved in the transcriptional regulation of genes required for mesoderm formation and differentiation. Binds to a palindromic site (called T site) and activates gene transcription when bound to such a site.
  • Involvement in disease

    Genetic variations in T are associated with susceptibility to neural tube defects (NTD) [MIM:182940]. NTD are common congenital malformations. Spina bifida, which results from malformations in the caudal region of the neural tube, is compatible with life but associated with significant morbidity, including lower limb paralysis.
    T is involved in susceptibility to the development of chordoma (CHDM) [MIM:215400]. Chordomas are rare, clinically malignant tumors derived from notochordal remnants. They occur along the length of the spinal axis, predominantly in the sphenooccipital, vertebral and sacrococcygeal regions. They are characterized by slow growth, local destruction of bone, extension into adjacent soft tissues and rarely, distant metastatic spread. Note=Susceptibility to development of chordomas is due to a T gene duplication.
  • Sequence similarities

    Contains 1 T-box DNA-binding domain.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession O15178 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6862 Human
    • Entrez Gene: 20997 Mouse
    • Omim: 601397 Human
    • SwissProt: O15178 Human
    • SwissProt: P20293 Mouse
    • Unigene: 389457 Human
    • Unigene: 913 Mouse
    • Alternative names

      • BRAC_HUMAN antibody
      • Brachyury homolog antibody
      • Brachyury protein antibody
      • Bry antibody
      • MGC104817 antibody
      • Protein T antibody
      • SAVA antibody
      • T antibody
      • T brachyury homolog antibody
      • T brachyury transcription factor antibody
      • T Protein antibody
      • T, brachyury homolog (mouse) antibody
      • TFT antibody
      • Transcription factor T antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-Brachyury / Bry antibody (ab20680)
      Immunocytochemistry/ Immunofluorescence - Anti-Brachyury / Bry antibody (ab20680)This image is courtesy of Ludovic Valllier, University of Cambridge, UK
      Anti-Brachyury antibody, ab20680, stained nuclei of human embryonic stem cells differentiated into mesendoderm. As would be expected, mesoderm cells express Brachyury whereas endoderm cells are negative.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody (ab20680)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody (ab20680)This image is courtesy of an Abreview submitted by Jim Manavis
      ab20680 staining Brachyury in Human brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% serum for 30 minutes at room temperature; antigen retrieval was by heat mediation with a citrate buffer. Samples were incubated with primary antibody (1/2000 in horse serum) for 12 hours. A Streptavidin-conjugated Horse anti-rabbit monoclonal (1/250) was used as the secondary antibody.

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody (ab20680)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody (ab20680)This image is courtesy of Amanda Evans, University of Cambridge
      A proportion of nuclei from Day 3 Mouse EBs (derived from R1 cells) stained positive for Brachury using ab20680. In Day 8 EBs a vastly reduced proportion of nuclei stained positive (not shown). In day 10 EBs no nuclei stained Brachyury-positive. The IHC-P data for Brachyury in EBs corresponded with qRT-PCR expression data (not shown). Controls on Day 3 EBs in which primary or secondary antibody were omitted, undifferentiated ES cells and feeder cells (MEFs) all exhibited very little background staining. Zinc was used for tissue fixation.
    • Western blot - Anti-Brachyury / Bry antibody (ab20680)
      Western blot - Anti-Brachyury / Bry antibody (ab20680)
      All lanes : Anti-Brachyury / Bry antibody (ab20680) at 1 µg/ml

      Lane 1 : MUG-Chor1 (human sacral bone chordoma) at 10 µg
      Lane 2 : MCF7 whole cell lysate (negative control) at 15 µg
      Lane 3 : Human embryonic stem cells (pluripotent) (negative control) at 10 µg
      Lane 4 : Human embryonic stem cells (mesoderm) at 10 µg

      Secondary
      All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 4 minutes


      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab20680 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    • Western blot - Anti-Brachyury / Bry antibody (ab20680)
      Western blot - Anti-Brachyury / Bry antibody (ab20680)
      Anti-Brachyury / Bry antibody (ab20680) at 1 µg/ml + Whole Cell Lysate from Murine ES Cells Differentiated to Express Mesoderm Markers at 15 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 53 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.



      ab20680 recognizes a band of 53 kDa in Murine Embryonic Stem Cells differentiated to express mesoderm markers. This corresponds in size to the Brachyury protein which has a predicted molecular weight of 47 kDa. A band of 75 kDa has also been observed, although we are unsure as to the identity of this band.

    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 (49)

    Publishing research using ab20680? Please let us know so that we can cite the reference in this datasheet.

    ab20680 has been referenced in 49 publications.

    • Ou M  et al. Single-cell sequencing reveals the potential oncogenic expression atlas of human iPSC-derived cardiomyocytes. Biol Open 10:N/A (2021). PubMed: 33589441
    • Chen M  et al. Transactivation of SOX5 by Brachyury promotes breast cancer bone metastasis. Carcinogenesis 41:551-560 (2020). PubMed: 31713604
    • Kim YY  et al. Effects of Natural Progesterone and Synthetic Progestin on Germ Layer Gene Expression in a Human Embryoid Body Model. Int J Mol Sci 21:N/A (2020). PubMed: 31991577
    • Xu J  et al. The Role of Transcriptional Factor Brachyury on Cell Cycle Regulation in Non-small Cell Lung Cancer. Front Oncol 10:1078 (2020). PubMed: 32719747
    • Wang L  et al. Establishment of a feeder and serum-free culture system for human embryonic stem cells. Zygote 28:175-182 (2020). PubMed: 31965957
    View all Publications for this product

    Customer reviews and Q&As

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

    1-6 of 6 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Brachyury / Bry antibody

    Inconclusive
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: RT°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate, EDTA, none
    Sample
    Zebrafish Tissue sections (Sagittal sections of whole formalin fixed, decalci)
    Specification
    Sagittal sections of whole formalin fixed, decalci
    Permeabilization
    No
    Fixative
    Formaldehyde
    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 Sep 19 2013

    Western blot abreview for Anti-Brachyury / Bry antibody - Mesoderm Marker

    Average
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse ES Cells)
    Loading amount
    50 µg
    Specification
    Mouse ES Cells
    Treatment
    Differentiating Conditions (Minus LIF Medium)
    Gel Running Conditions
    Reduced Denaturing (12)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    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.

    Mr. Utkarsh Kapoor

    Verified customer

    Submitted Dec 28 2012

    Western blot abreview for Anti-Brachyury / Bry antibody - Mesoderm Marker

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (6.5 embryo)
    Loading amount
    5 µg
    Specification
    6.5 embryo
    Gel Running Conditions
    Reduced Denaturing (9%)
    Blocking step
    Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    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 Aug 25 2011

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Brachyury / Bry antibody - Mesoderm Marker

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Brain)
    Specification
    Brain
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
    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. Jim Manavis

    Verified customer

    Submitted Oct 22 2010

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Brachyury / Bry antibody - Mesoderm Marker

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Embryoid bodies)
    Specification
    Embryoid bodies
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    None
    Permeabilization
    Yes - triton-x100
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
    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 May 28 2009

    Question


    Thanks for letting me know, that makes sense now!
    I wonder if I could ask you a favour, as I am unable to get the antibody
    to work could I ask, would it be possible to let me know the protocol you
    used to induce mesoderm please in the mouse embryonic stem cells? Was it
    spontaneous or was it a specific directed differentiation?
    If you have access to that info that'd be great as I could try it here and
    find out definitively if there is something strange going on!

    Read More

    Abcam community

    Verified customer

    Asked on Jul 03 2012

    Answer

    Thank you for contacting us.



    A. has forwarded your email to us in Scientific Support, and I would like to know what kind of application you are using?


    This way,we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


    I am looking forward to recieve your reply.


    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Jul 03 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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