Human/Mouse Brachyury Antibody AF2085: R&D Systems

Human/Mouse Brachyury Antibody

(32 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human Brachyury in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant human (rh) TBX-6, rhTBX-2, rhTBX-5, and rhTBX-18 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Brachyury
    Ser2-Glu202
    Accession # O15178
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Chromatin Immunoprecipitation (ChIP)
    Brachyury/DNA immunocomplexes were detected in sheared chromatin from fixed BG01V Human embryonic stem cells using standard PCR for the VEGF promoter sequence. Use 5 μg of antibody per 5x106 cells.
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Recombinant Human Brachyury by Western Blot. Western blot shows 10 ng of Recombinant Human Brachyury 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 (Catalog # 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.
Immunocytochemistry
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 Northern­Lights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counter­stained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
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 counter­stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
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 Northern­Lights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counter­stained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Chromatin Immunoprecipitation (ChIP)
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 (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The VEGF promoter was detected by standard PCR.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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.
  • Entrez Gene IDs:
    6862 (Human)
  • Alternate Names:
    brachyury protein; Brachyury; MGC104817; Protein T; T brachyury (mouse) homolog; T brachyury homolog; T, brachyury homolog (mouse); TF; TFT
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

32 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Guided self-organization and cortical plate formation in human brain organoids
    Authors: MA Lancaster, NS Corsini, S Wolfinger, EH Gustafson, AW Phillips, TR Burkard, T Otani, FJ Livesey, JA Knoblich
    Nat. Biotechnol., 2017;0(0):.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  2. CryoPause: A New Method to Immediately Initiate Experiments after Cryopreservation of Pluripotent Stem Cells
    Authors: KG Wong, SD Ryan, K Ramnarine, SA Rosen, SE Mann, A Kulick, E De Stanchi, FJ Müller, TJ Kacmarczyk, C Zhang, D Betel, MJ Tomishima
    Stem Cell Reports, 2017;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  3. In Vitro Micropatterned Human Pluripotent Stem Cell Test (�P-hPST) for Morphometric-Based Teratogen Screening
    Authors: J Xing, Y Cao, Y Yu, H Li, Z Song, H Yu
    Sci Rep, 2017;7(1):8491.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  4. Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells
    PLoS ONE, 2016;11(7):e0159246.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC - Fresh
  5. Dynamic Heterogeneity of Brachyury in Mouse Epiblast Stem Cells Mediates Distinct Response to Extrinsic Bone Morphogenetic Protein (BMP) Signaling
    Authors: L Song, J Chen, G Peng, K Tang, N Jing
    J. Biol. Chem., 2016;291(29):15212-25.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  6. Derivation of Patient Specific Pluripotent Stem Cells Using Clinically Discarded Cumulus Cells
    PLoS ONE, 2016;11(11):e0165715.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC - Paraffin embedded
  7. ROCK inhibitor primes human induced pluripotent stem cells to selectively differentiate towards mesendodermal lineage via epithelial-mesenchymal transition-like modulation
    Stem Cell Res, 2016;17(2):222-227.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC Not Specified
  8. Lineage tracing of neuromesodermal progenitors reveals novel Wnt-dependent roles in trunk progenitor cell maintenance and differentiation.
    Authors: Garriock R, Chalamalasetty R, Kennedy M, Canizales L, Lewandoski M, Yamaguchi T
    Development, 2015;142(9):1628-38.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Frozen
  9. An alternative pluripotent state confers interspecies chimaeric competency.
    Authors: Wu J, Okamura D, Li M, Suzuki K, Luo C, Ma L, He Y, Li Z, Benner C, Tamura I, Krause M, Nery J, Du T, Zhang Z, Hishida T, Takahashi Y, Aizawa E, Kim N, Lajara J, Guillen P, Campistol J, Esteban C, Ross P, Saghatelian A, Ren B, Ecker J, Izpisua Belmonte J
    Nature, 2015;521(7552):316-21.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC Not Specified
  10. Combinatorial analysis of developmental cues efficiently converts human pluripotent stem cells into multiple neuronal subtypes.
    Authors: Maury Y, Come J, Piskorowski R, Salah-Mohellibi N, Chevaleyre V, Peschanski M, Martinat C, Nedelec S
    Nat Biotechnol, 2015;33(1):89-96.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC OCT-embedded
  11. PI3K/mTORC2 regulates TGF-beta/Activin signalling by modulating Smad2/3 activity via linker phosphorylation.
    Authors: Yu J, Ramasamy T, Murphy N, Holt M, Czapiewski R, Wei S, Cui W
    Nat Commun, 2015;6(0):7212.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  12. SRF is essential for mesodermal cell migration during elongation of the embryonic body axis.
    Authors: Schwartz B, Marks M, Wittler L, Werber M, Wahrisch S, Nordheim A, Herrmann B, Grote P
    Mech Dev, 2014;133(0):23-35.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  13. Insulin-like growth factor promotes cardiac lineage induction in vitro by selective expansion of early mesoderm.
    Authors: Engels M, Rajarajan K, Feistritzer R, Sharma A, Nielsen U, Schalij M, de Vries A, Pijnappels D, Wu S
    Stem Cells, 2014;32(6):1493-502.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Flow
  14. Distinct Wnt-driven primitive streak-like populations reflect in vivo lineage precursors.
    Authors: Tsakiridis, Anestis, Huang, Yali, Blin, Guillaum, Skylaki, Stavroul, Wymeersch, Filip, Osorno, Rodrigo, Economou, Costas, Karagianni, Eleni, Zhao, Suling, Lowell, Sally, Wilson, Valerie
    Development, 2014;141(6):1209-21.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  15. Induction of pluripotency in human somatic cells via a transient state resembling primitive streak-like mesendoderm.
    Authors: Takahashi K, Tanabe K, Ohnuki M, Narita M, Sasaki A, Yamamoto M, Nakamura M, Sutou K, Osafune K, Yamanaka S
    Nat Commun, 2014;5(0):3678.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC - PFA fixed
  16. The role of Hath6, a newly identified shear-stress-responsive transcription factor, in endothelial cell differentiation and function.
    Authors: Fang F, Wasserman S, Torres-Vazquez J, Weinstein B, Cao F, Li Z, Wilson K, Yue W, Wu J, Xie X, Pei X
    J Cell Sci, 2014;127(0):1428-40.
    Species: Human
    Sample Type: Whole Cells
    Application: IF
  17. T-box transcription factor brachyury is associated with prostate cancer progression and aggressiveness.
    Authors: Pinto F, Pertega-Gomes N, Pereira M, Vizcaino J, Monteiro P, Henrique R, Baltazar F, Andrade R, Reis R
    Clin Cancer Res, 2014;20(18):4949-61.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  18. In vitro generation of neuromesodermal progenitors reveals distinct roles for wnt signalling in the specification of spinal cord and paraxial mesoderm identity.
    Authors: Gouti, Mina, Tsakiridis, Anestis, Wymeersch, Filip J, Huang, Yali, Kleinjung, Jens, Wilson, Valerie, Briscoe, James
    PLoS Biol, 2014;12(8):e1001937.
    Species: Human
    Sample Type: Whole Cells
    Application: IF
  19. Oct4 is required ~E7.5 for proliferation in the primitive streak.
    Authors: DeVeale B, Brokhman I, Mohseni P, Babak T, Yoon C, Lin A, Onishi K, Tomilin A, Pevny L, Zandstra P, Nagy A, van der Kooy D
    PLoS Genet, 2013;9(11):e1003957.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC
  20. The endothelial cell line bEnd.3 maintains human pluripotent stem cells.
    Authors: Joubin, Katherin, Richardson, Amelia, Novoa, Natalia, Tu, Edmund, Tomishima, Mark J
    Stem Cells Dev, 2012;21(12):2312-21.
    Species: Human
    Sample Type: Whole Cells
  21. Genomic targets of Brachyury (T) in differentiating mouse embryonic stem cells.
    Authors: Evans AL, Faial T, Gilchrist MJ
    PLoS ONE, 2012;7(3):e33346.
    Species: Human
    Sample Type: Cell Lysates
    Application: ChIP
  22. A novel chemically directed route for the generation of definitive endoderm from human embryonic stem cells based on inhibition of GSK-3.
    Authors: Bone HK, Nelson AS, Goldring CE, Tosh D, Welham MJ
    J. Cell. Sci., 2011;124(0):1992-2000.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  23. A late requirement for Wnt and FGF signaling during activin-induced formation of foregut endoderm from mouse embryonic stem cells.
    Authors: Hansson M, Olesen DR, Peterslund JM, Engberg N, Kahn M, Winzi M, Klein T, Maddox-Hyttel P, Serup P
    Dev. Biol., 2009;330(2):286-304.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  24. Noncanonical Wnt signaling orchestrates early developmental events toward hematopoietic cell fate from human embryonic stem cells.
    Authors: Vijayaragavan K, Szabo E, Bosse M, Ramos-Mejia V, Moon RT, Bhatia M
    Cell Stem Cell, 2009;4(3):248-62.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  25. TAZ controls Smad nucleocytoplasmic shuttling and regulates human embryonic stem-cell self-renewal.
    Authors: Varelas X, Sakuma R, Samavarchi-Tehrani P, Peerani R, Rao BM, Dembowy J, Yaffe MB, Zandstra PW, Wrana JL
    Nat. Cell Biol., 2008;10(7):837-48.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  26. Accelerated formation of multicellular 3-D structures by cell-to-cell cross-linking.
    Authors: De Bank PA, Hou Q, Warner RM, Wood IV, Ali BE, Macneil S, Kendall DA, Kellam B, Shakesheff KM, Buttery LD
    Biotechnol. Bioeng., 2007;97(6):1617-25.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  27. Creation of engineered human embryonic stem cell lines using phiC31 integrase.
    Authors: Thyagarajan B, Liu Y, Shin S, Lakshmipathy U, Scheyhing K, Xue H, Ellerstrom C, Strehl R, Hyllner J, Rao MS, Chesnut JD
    Stem Cells, 2007;26(1):119-26.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  28. A porous photocurable elastomer for cell encapsulation and culture.
    Authors: Gerecht S, Townsend SA, Pressler H, Zhu H, Nijst CL, Bruggeman JP, Nichol JW, Langer R
    Biomaterials, 2007;28(32):4826-35.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  29. Short-term BMP-4 treatment initiates mesoderm induction in human embryonic stem cells.
    Authors: Zhang P, Li J, Tan Z, Wang C, Liu T, Chen L, Yong J, Jiang W, Sun X, DU L, Ding M, Deng H
    Blood, 2007;111(4):1933-41.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  30. Activin a efficiently specifies definitive endoderm from human embryonic stem cells only when phosphatidylinositol 3-kinase signaling is suppressed.
    Authors: McLean AB, D'Amour KA, Jones KL, Krishnamoorthy M, Kulik MJ, Reynolds DM, Sheppard AM, Liu H, Xu Y, Baetge EE, Dalton S
    Stem Cells, 2007;25(1):29-38.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  31. Neurogenic effect of vascular endothelial growth factor during germ layer formation of human embryonic stem cells.
    Authors: Kim BK, Kim SE, Shim JH, Woo DH, Gil JE, Kim SK, Kim JH
    FEBS Lett., 2006;580(25):5869-74.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  32. Long-term self-renewal and directed differentiation of human embryonic stem cells in chemically defined conditions.
    Authors: Yao S, Chen S, Clark J, Hao E, Beattie GM, Hayek A, Ding S
    Proc. Natl. Acad. Sci. U.S.A., 2006;103(18):6907-12.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
Expand to show all 32 Citations
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Average Rating: 4 (Based on 1 review)

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We have 1 review tested in 1 species: Mouse.
We have 1 review tested in 1 application: Immunocytochemistry/Immunofluorescence.

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Immunocytochemistry/Immunofluorescence Human/Mouse Brachyury Antibody AF2085
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Very Good
 ICC/IF Mouse Anonymous 09/12/2016
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Immunocytochemistry/Immunofluorescence Human/Mouse Brachyury Antibody AF2085
Immunocytochemistry/Immunofluorescence: Human/Mouse Brachyury Antibody [AF2085]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedMesodermal cells differentiated from mouse ESCs,Embryonic stem cells
SpeciesMouse

Other Experimental Details

Other Experimental DetailsImmunohistochemistry 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).

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