Anti-TBR2 / Eomes antibody - ChIP Grade (ab23345)

Overview

  • Product name
    Anti-TBR2 / Eomes antibody - ChIP Grade
    See all TBR2 / Eomes primary antibodies
  • Description
    Rabbit polyclonal to TBR2 / Eomes - ChIP Grade
  • Tested applications
    Suitable for: IHC-FrFl, IHC-Fr, WB, ICC/IF, IHC-P, ChIP/Chip, ChIP, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Common marmoset
    Predicted to work with: Cow
  • Immunogen

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

    (Peptide available as ab25698.)

  • Positive control
    • WB: Human ES Cells treated with Retinoic Acid (24h) Whole Cell Lysate, Mouse Embryonic Brain (E14) Tissue Lysate. IHC-Fr: Mouse Embryonic Brain (E14.5).
  • General notes

    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.

Properties

Applications

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.

Application Abreviews Notes
IHC-FrFl Use at an assay dependent concentration.
IHC-Fr 1/500.

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

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.

Target

  • Function
    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.
  • Tissue specificity
    Expressed in CD8+ T-cells.
  • Involvement in disease
    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.
  • Sequence similarities
    Contains 1 T-box DNA-binding domain.
  • Developmental stage
    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.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Anti-TBR2 / Eomes antibody - ChIP Grade images

  • 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.

  • 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.

  • 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
    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

  • 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

  • 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
    Lane 3 : EL4 cells + V5 tagged vector
    Lane 4 : EL4 cells + empty vector
    Lane 5 : EL4 cells + vector expressing V5 tagged Eomesodermin
    Lane 6 : EL4 cells + V5 tagged vector

    Secondary
    Goat anti Rabbit at 1/2500 dilution

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

    This image was submitted as part of a review published on 9th May 2006

    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
    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

    This image is a courtesy of Anonymous Abreview

    See Abreview

References for Anti-TBR2 / Eomes antibody - ChIP Grade (ab23345)

This product has been referenced in:
  • Subramanian L  et al. Dynamic behaviour of human neuroepithelial cells in the developing forebrain. Nat Commun 8:14167 (2017). IF ; Human . Read more (PubMed: 28139695) »
  • Subashini C  et al. Wnt5a is a crucial regulator of neurogenesis during cerebellum development. Sci Rep 7:42523 (2017). IHC-P ; Mouse . Read more (PubMed: 28205531) »

See all 122 Publications for this product

Product Wall

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
Username

Dr. Kate Hawkins

Verified customer

Submitted 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
Username

Naor Sagy

Verified customer

Submitted 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
Username

Ms. Xiaofeng Zhou

Verified customer

Submitted 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
Username

Abcam user community

Verified customer

Submitted 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
Username

Abcam user community

Verified customer

Submitted 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
Username

Abcam user community

Verified customer

Submitted May 04 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.3% Triton x 100: 30 min
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Dr. Md. Golam Sharoar

Verified customer

Submitted May 04 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (cortical neurons)
Permeabilization
Yes - 0.25% Triton X-100 in PBS
Specification
cortical neurons
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 07 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (H9 embryonic stem cells)
Specification
H9 embryonic stem cells
Permeabilization
Yes - 0.01% Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 22 2014

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: RT°C
Antigen retrieval step
None
Sample
Ferret Tissue sections (brain)
Specification
brain
Permeabilization
Yes - 0.5% Triton X 100 / PBS
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 26 2014

1-10 of 62 Abreviews or Q&A

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