Overview

  • Product nameAnti-BCL9 antibody
    See all BCL9 primary antibodies
  • Description
    Rabbit polyclonal to BCL9
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Human BCL9.

    (Peptide available as ab37304.)

  • Positive control
    • This antibody gave a positive signal in the following cell lysates: HeLa (Human epithelial carcinoma cell line) Nuclear Lysate; Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate; MCF7 (Human breast adenocarcinoma cell line) Nuclear Lysate; Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab37305 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
WB Use a concentration of 2 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 140 kDa).Can be blocked with Human BCL9 peptide (ab37304).
IHC-P Use at an assay dependent concentration. PubMed: 22914623
ICC/IF Use a concentration of 1 µg/ml.

Target

Anti-BCL9 antibody images

  • ICC/IF image of ab37305 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab37305, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal 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).

  • All lanes : Anti-BCL9 antibody (ab37305) at 2 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 2 : Jurkat nuclear extract lysate (ab14844)
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Nuclear Lysate
    Lane 4 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate

    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 : 140 kDa
    Observed band size : 180 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,47 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
  • Immunohistochemical analysis of Human colorectal cancer tissue, staining BCL9 with ab37305 at 5 µg/ml.

    Left panel: BCL9 antibody alone. Right panel: BCL9 antibody + ab37304 - BCL9 peptide.

References for Anti-BCL9 antibody (ab37305)

This product has been referenced in:
  • Zhao JJ  et al. miR-30-5p functions as a tumor suppressor and novel therapeutic tool by targeting the oncogenic Wnt/ß-catenin/BCL9 pathway. Cancer Res 74:1801-13 (2014). Human . Read more (PubMed: 24599134) »
  • Hyeon J  et al. Prognostic Significance of BCL9 Expression in Hepatocellular Carcinoma. Korean J Pathol 47:130-6 (2013). IHC-P ; Human . Read more (PubMed: 23667372) »

See all 4 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step Heat mediated
Sample Rhesus monkey Tissue sections (Colon)
Specification Colon
Permeabilization Yes - 0.1X Triton-X-100
Fixative Z-fix
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Submitted Dec 11 2014

Abreviews
Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (gel 12%)
Sample Human Cell lysate - whole cell (muscle)
Specification muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Oct 30 2014

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We do not know what effect H2O2 stress will have on expression of BCL9 in MCF7 cells therefore I would recommend perhaps running H2O2 treated MCF7 lysate alongside untreated if possible to determine affect. Read More

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