Recombinant
RabMAb

Recombinant Anti-Brachyury / Bry antibody [EPR18113] (ab209665)

Rabbit recombinant monoclonal Brachyury / Bry antibody [EPR18113]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 3 publication(s).

Overview

  • Product name
    Anti-Brachyury / Bry antibody [EPR18113]
    See all Brachyury / Bry primary antibodies
  • Description
    Rabbit monoclonal [EPR18113] to Brachyury / Bry
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, IP, WB, IHC-P, IHC - Wholemountmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Non human primates, Common marmoset
  • Immunogen

    Recombinant fragment within Human Brachyury/ Bry aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: O15178

  • Positive control
    • IHC-P: Human chordoma tissue and mouse E14.5 embryo tissue.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209665 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 1/1000.
Flow Cyt 1/400.
IP 1/1000.
WB 1/1000. Predicted molecular weight: 47 kDa.
IHC-P 1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC - Wholemount 1/100.

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.
  • Information by UniProt
  • Database links
  • 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

  • Ab209665 staining Brachyury/Bry in MUG-Chor1 by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were incubated with primary antibody at a 1/1000 dilution.  An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at a 1/1000 dilution.  An Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as a counterstain antibody.  DAPI was used as nuclear counterstain. Nuclear staining on MUG-Chor1 cells.

  • ab209665 staining Brachyury/Bry in mouse E14.5 embryo tissue sections by Immunohistochemistry (IHC-P). Tissue was fixed paraffin and antigen retrieval was performed by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at a 1/4000 dilution. A ready to use Goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter-stain. Nuclear staining on notochord of mouse E14.5 embryo.

  • Ab209665 staining Brachyury/Bry in Rat E14.5 embryo tissue sections by Immunohistochemistry (IHC-P). Tissue was fixed paraffin and antigen retrieval was performed by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at a 1/4000 dilution. A ready to use Goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter-stain. Nuclear staining on notochord of rat E14.5 embryo.

  • IHC - Wholemount of E6.5 wholemount mouse embryo labelling Brachyury / Bry with ab209665. Sample was incubated with primary antibody (1/100 in PBS/0.1% Triton-X/5% donkey serum/1% BSA) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG monoclonal was used as the secondary antibody (1/500).

    See Abreview

  • Immunohistochemical analysis of paraffin-embedded Human chordoma tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Human chordoma is observed. Counter stained with Hematoxylin

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Lane 1 (input):  MUG-Chor1 (human sacral bone chordoma) whole cell lysate, 10 μg
    Lane 2 (+): MUG-Chor1 whole cell lysate
    Lane 3 (-):  Rabbit monoclonal IgG (ab172730) instead of  ab209665 in MUG-Chor1 whole cell lysate

    Ab209665 immunoprecipitating Brachyury/Bry in MUG-Chor1 lysates. For western blotting, primary antibody used was ab209665 at 1/1000 dilution.  VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking and diluting buffer used was 5% NFDM/TBST.

  • Ab209665 staining Brachyury/Bry in MUG-Chor1 (Human sacral bone chordoma) cell line by Flow cytometry. Cells were fixed with 4% paraformaldehyde and 90% methanol. Sample was incubated with primary antibody at 1/400 dilution (red). A Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution.  Rabbit monoclonal IgG (ab172730) (Black) was used as an isotype control. Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human kidney. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Anti-Brachyury / Bry antibody [EPR18113] (ab209665) at 1/1000 dilution + MUG-Chor1 (human sacral bone chordoma), whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 47 kDa
    Observed band size: 43,49 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded Human meningioma tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human meningioma. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human chondrosarcoma tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human chondrosarcoma. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

     

References

This product has been referenced in:
  • Frith TJR & Tsakiridis A Efficient Generation of Trunk Neural Crest and Sympathetic Neurons from Human Pluripotent Stem Cells Via a Neuromesodermal Axial Progenitor Intermediate. Curr Protoc Stem Cell Biol N/A:e81 (2019). Read more (PubMed: 30688409) »
  • Frith TJ  et al. Human axial progenitors generate trunk neural crest cells in vitro. Elife 7:N/A (2018). Read more (PubMed: 30095409) »
See all 4 Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Differentiated iPS cells)
Permeabilization
Yes - 0.3% Triton + 0.1% BSA
Specification
Differentiated iPS cells
Blocking step
0.1% BSA/10%FCS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 03 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Differentiated Cell)
Permeabilization
Yes - 0.5% Triton X-100
Specification
Differentiated Cell
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 05 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Mesoderm cells derived from hESCs)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
10 µg
Treatment
8uM CHIR99021
Specification
Mesoderm cells derived from hESCs
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 03 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Differentiated Embryonic Stem Cells)
Permeabilization
Yes - 0.5% Triton X-100
Specification
Differentiated Embryonic Stem Cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Mr. Dylan Stavish

Verified customer

Submitted Apr 26 2018

Application
IHC - Wholemount
Sample
Mouse Embryo (E6.5 wholemount mouse embryo)
Specification
E6.5 wholemount mouse embryo

Dr. Sophie Morgani

Verified customer

Submitted Feb 09 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jan 16 2017

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