Overview

  • Product name
  • Description
    Rabbit polyclonal to TLE3
  • Tested applications
    Suitable for: IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 to the C-terminus of Human TLE3.

    (Peptide available as ab109092.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: Breast (Human) Tissue Lysate; MCF7 Whole Cell Lysate; MDA-MB-231 Whole Cell Lysate; Lung (Human) Tissue Lysate; A549 Whole Cell Lysate; A431 Whole Cell Lysate; HeLa Nuclear Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab94972 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
IP Use at an assay dependent concentration.
WB Use a concentration of 1 mg/ml. Detects a band of approximately 83 kDa (predicted molecular weight: 83 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Transcriptional corepressor that binds to a number of transcription factors. Inhibits the transcriptional activation mediated by CTNNB1 and TCF family members in Wnt signaling. The effects of full-length TLE family members may be modulated by association with dominant-negative AES.
  • Tissue specificity
    Placenta and lung.
  • Sequence similarities
    Belongs to the WD repeat Groucho/TLE family.
    Contains 7 WD repeats.
  • Post-translational
    modifications
    Ubiquitinated by XIAP/BIRC4. This ubiquitination does not affect its stability, nuclear localization, or capacity to tetramerize but inhibits its interaction with TCF7L2/TCF4.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • E(sp1) homolog Drosophila antibody
    • Enhancer of split groucho 3 antibody
    • Enhancer of split groucho-like protein 3 antibody
    • ESG 3 antibody
    • ESG antibody
    • ESG3 antibody
    • FLJ39460 antibody
    • GRG 3 antibody
    • GRG3 antibody
    • HsT18976 antibody
    • KIAA1547 antibody
    • TLE 3 antibody
    • Tle3 antibody
    • TLE3_HUMAN antibody
    • Transducin like enhancer of split 3 antibody
    • Transducin like enhancer of split 3 homolog of Drosophila E(sp1) antibody
    • Transducin like enhancer protein 3 antibody
    • transducin-like enhancer of split 3 (E(sp1) homolog, Drosophila) antibody
    • Transducin-like enhancer protein 3 antibody
    see all

Images

  • TLE3 was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Rabbit polyclonal to TLE3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab94972.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 83kDa; TLE3

  • All lanes : Anti-TLE3 antibody (ab94972) at 1 µg/ml

    Lane 1 : Human breast tissue lysate - total protein (ab30090)
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : Lung (Human) Tissue Lysate
    Lane 5 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
    Lane 6 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 7 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 83 kDa
    Observed band size : 83 kDa
    Additional bands at : 125 kDa,43 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutesAbcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
  • ICC/IF image of ab94972 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab94972, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml.

References

This product has been referenced in:
  • Yang RW  et al. TLE3 represses colorectal cancer proliferation by inhibiting MAPK and AKT signaling pathways. J Exp Clin Cancer Res 35:152 (2016). Read more (PubMed: 27669982) »
  • Bara AM  et al. Generation of a TLE3 heterozygous knockout human embryonic stem cell line using CRISPR-Cas9. Stem Cell Res 17:441-443 (2016). Read more (PubMed: 27879221) »

See all 2 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
30 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Submitted Oct 23 2017

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