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

Overview

  • Product nameAnti-TBR2 / Eomes antibody - ChIP Grade
    See all TBR2 / Eomes primary antibodies
  • Description
    Rabbit polyclonal to TBR2 / Eomes - ChIP Grade
  • Tested applicationsSuitable for: IHC-FrFl, IHC-Fr, WB, ICC/IF, IHC-P, ChIP/Chip, ChIP, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Marmoset (common)
    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
    • Human Mesendoderm (Day 2) Whole Cell Lysate, Mouse Embryonic Brain (E14) Tissue Lysate
  • 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.
WB Use a concentration of 0.4 - 2.5 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 72 kDa).
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

  • FunctionFunctions 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 specificityExpressed in CD8+ T-cells.
  • Involvement in diseaseNote=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 similaritiesContains 1 T-box DNA-binding domain.
  • Developmental stageDetected 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 localizationNucleus.
  • 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

  • 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
  • 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.

  • 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

  • ICC/IF image of ab23345 stained human SH-SY5Y cells. The cells were PFA fixed (10 min), permeabilised in TBS-T (20 min) and incubated with the antibody (ab23345, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • 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.

  • 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

  • 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.
  • 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 1/1000 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:

See all 100 Publications for this product

Product Wall

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
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Abcam user community

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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
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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
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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
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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
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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
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Submitted Jun 26 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 23°C
Sample Human Cell (Radial glial cells differentiated from human embry)
Specification Radial glial cells differentiated from human embry
Permeabilization Yes - 0.3% Triton X-100
Fixative Paraformaldehyde
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Submitted Nov 11 2013

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer ph 6.0
Sample Rat Tissue sections (Adult rat olfactory bulb)
Specification Adult rat olfactory bulb
Permeabilization Yes - 0.3% Triton X-100
Fixative Paraformaldehyde
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Submitted Oct 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step None
Sample Mouse Tissue sections (E15 mouse cortex)
Specification E15 mouse cortex
Permeabilization Yes - 0.1% Triton X-100
Fixative Paraformaldehyde
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Submitted Oct 01 2013

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