製品の概要

  • 製品名
    Anti-TBR2 / Eomes antibody - ChIP Grade
    TBR2 / Eomes 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to TBR2 / Eomes - ChIP Grade
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: IHC-FrFl, IHC-Fr, WB, ICC/IF, IHC-P, ChIP/Chip, ChIP, IHC-FoFrmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Common marmoset
    交差が予測される動物種: Cow
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 650 to the C-terminus of Mouse TBR2/ Eomes.

    Immunogen の所有権に関して (Peptide available as ab25698.)

  • ポジティブ・コントロール
    • WB: Human ES Cells treated with Retinoic Acid (24h) Whole Cell Lysate, Mouse Embryonic Brain (E14) Tissue Lysate; Human Mesendoderm (Day 2) Whole Cell Lysate; EL4 cells + vector expressing V5 tagged Eomesodermin. IHC-Fr: Mouse Embryonic Brain (E14.5); Adult C57/BL6 mouse forebrain coronal sections; Mouse developing cerebral cortex tissue; Mouse embryonic brain tissue. ICC/IF: Human embryonic stem cells differentiated into mesendoderm. IHC-P: Rat brain (e16) tissue.
  • 特記事項

    Tbr2 expression is observed in neuron progenitor compartments in development (the subventricular zone and ventricular zone) and expression rises and falls with cortical plate neurogenesis. Transition from radial glia to intermediate progenitor cell is associated with upregulation of Tbr2.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab23345 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-FrFl Use at an assay dependent concentration.
IHC-Fr 1/100 - 1/500.

Abcam recommends the following antigen retrieval method: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).

We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody.

WB Use a concentration of 0.4 - 2.5 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 72 kDa).

Abcam recommends using milk as the blocking agent.

ICC/IF Use at an assay dependent concentration.
IHC-P 1/500.
ChIP/Chip Use at an assay dependent concentration. PubMed: 20176728
ChIP Use at an assay dependent concentration.
IHC-FoFr Use a concentration of 1 - 2 µg/ml.

ターゲット情報

  • 機能
    Functions as a transcriptional activator playing a crucial role during development. Functions in trophoblast differentiation and later in gastrulation, regulating both mesoderm delamination and endoderm specification. Plays a role in brain development being required for the specification and the proliferation of the intermediate progenitor cells and their progeny in the cerebral cortex. Also involved in the differentiation of CD8+ T-cells during immune response regulating the expression of lytic effector genes.
  • 組織特異性
    Expressed in CD8+ T-cells.
  • 関連疾患
    Note=A translocation t(3;10)(p24;q23) located 215 kb 3' to the EOMES gene but leading to loss of its expression was identified in a large consanguineous family. Homozygous silencing produces microcephaly associated with corpus callosum agenesis, bilateral polymicrogyria, ventricular dilatation and a small cerebellum.
  • 配列類似性
    Contains 1 T-box DNA-binding domain.
  • 発生段階
    Detected at 7 weeks of development in the forebrain floorplate of the CNS. Expressed within the mantle layer and migrating neuroblasts of the telencephalon at 12.5 weeks of development.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • eomes antibody
    • EOMES_HUMAN antibody
    • Eomesodermin antibody
    • Eomesodermin homolog antibody
    • t box brain 2 antibody
    • T-box brain protein 2 antibody
    • T-brain-2 antibody
    • Tbr 2 antibody
    • TBR-2 antibody
    see all

画像

  • Dlx2 and Tbr2 identify non overlapping progenitor lineages in the adult mouse SVZ (subventricular zone).

    (A–B) Adult C57/BL6 mouse forebrain coronal sections at rostro-caudal point 1.2 relative to the bregma were immunostained for Dcx (red), Dlx2 or Tbr2 (green) and Ki67 (blue). Both progenitor populations show characteristics of migrating neuroblasts, as indicated by their Dcx expression (C–D). Adult Mash1 mouse forebrain coronal sections at rostro-caudal point 1.2 relative to the bregma were immunostained for Tbr2 (red) and Dlx2 (blue). Both Tbr2 and Dlx2 exhibited EGFP expression, but showed no colocalisation. Right side captions show cropped individual channels and the merges. Full panel insets are zoomed and cropped DAB stained photomicrographs of rostral periventricular sections for Tbr2 in the dorsal SVZ (A), Dlx2 in the dorso-lateral SVZ (B) and Mash1 in the ventro-lateral SVZ (C). Yellow arrows and arrowheads show respectively positive stained cell and low level TF staining. Dotted lines mark approximate boundaries of ventricular space. Flattened confocal z-stacks are of 14–15 µm thickness, including captions. Scale bars: 15 µm in full panels, 20 µm in captions and 25 µm in insets.

  • Antigen retrieval was carried out by microwaving sections in 0.01M sodium citrate (pH6.0) for 20 minutes. Sections were washed with PBS-0.01% Triton X-100 and then incubated for 30 minutes in blocking solution containing 20% goat or donkey serum, then incubated overnight at 4°C with ab23345 (1:100). Following incubation with ab23345, sections were washed in PBS-0.01% Triton X-100 and incubated with the secondary antibody donkey anti-rabbit Alexa Fluor 568 for 1 hour at room temperature. Nuclei were counterstained with TOPRO-3 (1:1000). Fluorescent images were captured using a Leica NTS confocal microscope.

  • Paraffin-embedded rat brain (e16) tissue stained for TBR2 / Eomes using ab23345 at 1/10000 dilution in immunohistochemical analysis, followed by anti Rabbit IgG Biotin.

    See Abreview

  • Human embryonic stem cells were differentiated into mesendoderm. Cells were then stained with DAPI (blue) and a TBR2 / Eomes specific antibody, ab23345 (1:100, red).

    Ludovic Vallier, University of Cambridge

  • Anti-TBR2 / Eomes antibody - ChIP Grade (ab23345) at 1 µg/ml + Human ES Cells treated with Retinoic Acid (24h) at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 72 kDa
    Observed band size: 85 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.


    Exposure time: 20 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 3% Milk before being incubated with ab23345 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    Abcam recommends using milk as the blocking agent.

  • IHC image of TRB2 staining in a mouse brain E14 frozen tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6). Non-specific protein-protein interactions were then blocked in TBS containing 0.2% (v/v) Triton X-100 for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.05% (v/v) Triton X-100 and 1% (w/v) BSA with ab23345 at 1/100 dilution. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody was used to detect the primary antibody.  The section was mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-TBR2 / Eomes antibody - ChIP Grade (ab23345) at 1 µg/ml (BLOCKED WITH 3% MILK)

    Lane 1 : Human Mesendoderm (Day 2) Whole Cell Lysate
    Lane 2 : E14 Mouse Embryo Brain Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 72 kDa
    Observed band size: 85 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.


    Exposure time: 4 minutes
  • ab23345 staining mouse developing cerebral cortex tissue sections by IHC-Fr.  Sections were PFA fixed and permeabilized in TX-100 prior to blocking with 2.5% serum for 1 hour at RT.  The primary antibody was diluted 1/500 and incubated with the sample for 18 hours.  A biotinylated pig anti-rabbit IgG antibody, diluted 1/500, was used as the secondary.

    See Abreview

  • ab23345 staining TBR2 / Eomes in mouse embryonic brain tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with formaldehyde and blocking with 1% BSA and normal Goat serum for 30 minutes at RT. The sample was incubated with primary antibody (1/1000 in TBS + BSA 1%) for 10 hours at 40C. An Alexa Fluor® 555-conjugated Goat polyclonal to rabbit IgG was used as secondary antibody at 1/800 dilution.

    See Abreview

  • Lanes 1-3 : Anti-TBR2 / Eomes antibody - ChIP Grade (ab23345) at 1/2000 dilution
    Lanes 4-6 : V5 antibody

    Lane 1 : EL4 cells + empty vector
    Lane 2 : EL4 cells + vector expressing V5 tagged Eomesodermin
    Lanes 3 & 6 : EL4 cells + V5 tagged vector
    Lane 4 : EL4 cells + empty vector
    Lane 5 : EL4 cells + vector expressing V5 tagged Eomesodermin

    Secondary
    All lanes : Goat anti Rabbit at 1/2500 dilution

    Predicted band size: 72 kDa
    Observed band size: 72 kDa



    ab23345 detects a clear band of ~ 72 kDa in lysates from EL4 cells expressing V5 tagged Eomesodermin (lane 2).  Lanes 1 and 3 contain lysates from EL4 cells expressing empty vector or V5 tag alone. Lanes 4-6 show the same lysates blotted with anti-V5 tag antibody. GAPDH was used as a loading control.

  • All lanes : Anti-TBR2 / Eomes antibody - ChIP Grade (ab23345) at 1 µg/ml

    Lane 1 : Human Mesendoderm (Day 2) Whole Cell Lysate
    Lane 2 : Human Mesendoderm (Day 2) Whole Cell Lysate with Mouse TBR2 / Eomes peptide (ab25698) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size: 72 kDa
    Observed band size: 85 kDa why is the actual band size different from the predicted?
    Additional bands at: 74 kDa. We are unsure as to the identity of these extra bands.

  • Anti-TBR2 / Eomes antibody - ChIP Grade (ab23345) at 1/1000 dilution + Lysate prepared from mouse embryonic brain tissue at 20 µg

    Secondary
    HRP-conjugated goat polyclonal to rabbit IgG

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 72 kDa
    Observed band size: 72 kDa


    Exposure time: 5 minutes

    See Abreview

参考文献

This product has been referenced in:
  • Pfeiffer MJ  et al. Cardiogenic programming of human pluripotent stem cells by dose-controlled activation of EOMES. Nat Commun 9:440 (2018). Read more (PubMed: 29382828) »
  • Ziegler-Waldkirch S  et al. Seed-induced Aß deposition is modulated by microglia under environmental enrichment in a mouse model of Alzheimer's disease. EMBO J 37:167-182 (2018). IHC-Fr ; Mouse . Read more (PubMed: 29229786) »
See all 223 Publications for this product

レビューと Q&A

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1-10 of 38 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Common marmoset Tissue sections (Cerebellum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Cerebellum
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

投稿 Jul 27 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (e16 brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Specification
e16 brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

投稿 May 18 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (e14/15 brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Specification
e14/15 brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

投稿 May 03 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (E13 brain tissue)
Permeabilization
Yes - 0.3% Triton X-100
Specification
E13 brain tissue
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde

Dr. Bhavin Shah

Verified customer

投稿 Feb 27 2018

Application
Immunocytochemistry
Sample
Human Cell (Neural precursor cells)
Permeabilization
Yes - 0.3% triton
Specification
Neural precursor cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Kate Hawkins

Verified customer

投稿 Aug 03 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (ESC?)
Permeabilization
No
Specification
ESC?
Blocking step
Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde

Naor Sagy

Verified customer

投稿 Jul 03 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (E14.5 forebrain)
Gel Running Conditions
Non-reduced Denaturing (4-20% TGX)
Loading amount
30 µg
Specification
E14.5 forebrain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Ms. Xiaofeng Zhou

Verified customer

投稿 May 22 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (Kidney)
Permeabilization
Yes - 0.1% troton x-100
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

投稿 Jun 20 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Brain)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

投稿 May 04 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Monkey Cell lysate - whole cell (Kidney)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

投稿 May 04 2016

1-10 of 38 Abreviews

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