Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Rat, Avian, Avian - Chicken, Chicken, Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque), Transgenic Mouse, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation Sequencing, In vivo assay, IF/ICC, Immunolocalization

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Brachyury Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Brachyury
Ser2-Glu202
Accession # O15178

Specificity

Detects human Brachyury in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse Brachyury Antibody

Detection of Recombinant Human Brachyury antibody by Western Blot.

Detection of Recombinant Human Brachyury by Western Blot.

Western blot shows 10 ng of Recombinant Human Brachyury (amino acids Ser2-Glu202) and Recombinant Human TBX6. PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for Brachyury at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Detection of Human Brachyury by Western Blot.

Western blot shows lysates of NCI‑H460 human large cell lung carcinoma cell line and MCF 10A human breast epithelial cell line (negative control). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Brachyury at approximately 50 kDa (as indicated). GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Brachyury antibody in Differentiated Human Embryonic Stem Cells by Immunocytochemistry (ICC).

Brachyury in Differentiated Human Embryonic Stem Cells.

Brachyury was detected in immersion fixed differentiated human embryonic stem cells using 10 µg/mL Goat Anti-Human Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) for 3 hours at room temperature. Cells were stained (green) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Brachyury antibody in BG01V Human Stem Cells by Immunocytochemistry (ICC).

Brachyury in BG01V Human Stem Cells.

Brachyury was detected in immersion fixed BG01V human embryonic stem cells differentiated into mesoderm using Goat Anti-Human Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counter-stained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Brachyury antibody in Embryonic Mouse Notochord by Immunohistochemistry (IHC-Fr).

Brachyury in Embryonic Mouse Notochord.

Brachyury was detected in immersion fixed frozen sections of embryonic mouse notochord (E9.5) using 10 µg/mL Goat Anti-Human Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) overnight at 4 °C. Tissue was stained with the Northern-Lights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counter-stained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of Brachyury-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of Brachyury-regulated Genes by Chromatin Immunoprecipitation.

Mesoderm-differentiated BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Brachyury/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) or control antibody (AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (MAG999) and DNA was purified using chelating resin solution. TheVEGFpromoter was detected by standard PCR.

Detection of Brachyury in NCI-H460 cells by Flow Cytometry.

NCI-H460 cells were stained with Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Detection of Human Brachyury by Immunocytochemistry/Immunofluorescence

Detection of Human Brachyury by Immunocytochemistry/Immunofluorescence

Generation and characterization of systemic lupus erythematosus (SLE)‐specific human‐induced pluripotent stem cells (hiPSCs). (A) Dermal fibroblasts derived from patient with SLE were reprogrammed into iPSCs using Sendai virus vectors and three clones (#1, #2 and #3) were characterized. RT‐PCR confirms the loss of transgenes in hiPSCs‐SLE (lanes 1, 2 and 3), presence (lane 4) in infected fibroblasts (ipF‐SLE) and absence of Sendai viral transgenes in parental fibroblasts (pF‐SLE) (lane 5). Full‐length gels are presented in File S2. (B) The hiPSCs‐SLE colonies expressed alkaline phosphatase. Scale bar, 500 μm. (C) RT‐qPCR analysis of pluripotency genes OCT4, NANOG, SOX2 and REX1 was performed in fibroblasts and in hiPSCs derived from patient with SLE. All expression values are normalized to GAPDH and relative donor fibroblasts. Data are mean ± SEM and all statistical analysis was made between hiPSCs‐SLE clones and relative fibroblasts by Student's t test showing P‐values ≤ .05 in each comparison. (D) PluriTest assays combines novelty score (blue) on x‐axis and pluripotency score (red) on y‐axis. hiPSCs‐SLE localize in the red cloud suggesting the empirical distribution of pluripotent cells compared to non‐pluripotent blue cloud. (E) Representative images of M‐FISH staining show normal karyotypes of hiPSCs‐SLE clones. (F) Immunofluorescence analysis of pluripotent stem cell markers Nanog (green), Oct4 (red) and co‐staining with DAPI (blue) in hiPSCs‐SLE. Scale bar, 50 μm. (G) Representative images of floating and adherent EBs derived from hiPSCs‐SLE at differentiation day 8 and 18, respectively. Scale bar, 500 μm. (H) RT‐qPCR results confirm the capability of hiPSCs‐SLE to differentiate into all three germ layers. The expression levels of GATA4, HAND1 and PAX6 in EBs are relative to undifferentiated hiPSCs. All expression values are normalized to GAPDH and relative hiPSCs. Data are mean ± SEM and all statistical analysis was made between EBs‐SLE and relative hiPSCs‐SLE clones by Student's t test showing P‐values ≤ .05 in each comparison. (I) Immunostaining of Otx2 (ectoderm marker), Sox17 (endoderm marker), Brachyury (mesoderm marker) and co‐staining with DAPI (blue) in EBs derived from hiPSCs‐SLE. Scale bar, 50 μm Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31536674), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Brachyury by Immunocytochemistry/ Immunofluorescence

Detection of Mouse Brachyury by Immunocytochemistry/ Immunofluorescence

Generation of NMPs from EpiSCs.(A) Brachyury/Sox2 immunocytochemistry in EpiSC cultures treated with FGF/CHIR for 72 h. (B) qPCR analysis for indicated markers in mouse EpiSCs treated with FGF/CHIR. Error bars = s.d. (n = 3). n/d, not determined. Results are represented as log10 ratio of expression versus untreated EpiSCs. The data used to generate the plot can be found in Data S4. (C) Combined fluorescence/brightfield microscopy showing donor cell incorporation of grafted GFP+ EpiSC differentiated for 48 h in FGF/CHIR after 48 h embryo culture. (D) Table summarizing the incorporation of grafted GFP+ EpiSC differentiated for 24 h or 48 h in Fgf/Wnt within host embryos. NT, neural tube; Som, somite; PSM, presomitic mesoderm; n/a, not applicable. (E) Representative examples of donor cell incorporation (green, GFP) and differentiation (red, immunofluorescence for indicated markers). Cell nuclei were stained with DAPI (blue). White boxes indicate the position of magnified images of GFP+ cells. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157815), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse Brachyury Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed differentiated human embryonic stem cells and BG01V human embryonic stem cells differentiated into mesoderm

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed frozen sections of embryonic mouse notochord (E9.5)

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: NCI-H460 cells fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). 

Western Blot

0.1-1 µg/mL
Sample: Recombinant Human Brachyury and NCI‑H460 human large cell lung carcinoma cell line

Reviewed Applications

Read 4 reviews rated 4.3 using AF2085 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Brachyury

Brachyury is the founding member of the T-box family of transcription factors, which is characterized by the N-terminal conserved DNA-binding T-domain. Brachyury is required in the early determination and differentiation of mesoderms. Human brachyury molecule shares 90% homology with mouse brachyury.

Alternate Names

brachyury protein, MGC104817, Protein T, SAVA, T brachyury (mouse) homolog, T brachyury homolog, T Brachyury Transcription Factor, T, brachyury homolog (mouse), T-Box Transcription Factor T, TBXT, TF, TFT

Entrez Gene IDs

6862 (Human)

Gene Symbol

TBXT

UniProt

O15178

Additional Brachyury Products

Product Documents for Human/Mouse Brachyury Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse Brachyury Antibody

BG01V cells are licensed from ViaCyte, Inc.

For research use only

Citations for Human/Mouse Brachyury Antibody

Customer Reviews for Human/Mouse Brachyury Antibody (4)

4.3 out of 5
4 Customer Ratings
5 Stars
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4 Stars
25%
3 Stars
25%
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1 Stars
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Human/Mouse Brachyury Antibody
    Name: Leslie Priddy
    Application: Immunohistochemistry
    Sample Tested: Tumor cell lyastes
    Species: Mouse
    Verified Customer | Posted 04/13/2018
  • Human/Mouse Brachyury Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Mesodermal cells differentiated from mouse ESCs and Embryonic stem cells
    Species: Mouse
    Verified Customer | Posted 09/12/2016
    Immunohistochemistry on fixed 2D cell culture. Primary antibody was used at 5ug/mL and incubated overnight. Brachyury-positive cells are visualized in red, nuclei are counterstained by Dapi (blue).
    Human/Mouse Brachyury Antibody AF2085
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 24319666
    Species: Mouse
    Verified Customer | Posted 01/06/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 23658023
    Species: Human
    Verified Customer | Posted 01/06/2015

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