Overview

  • Product name

    Anti-BCL9 antibody
  • Description

    Rabbit polyclonal to BCL9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide corresponding to Human BCL9 aa 800-900 conjugated to keyhole limpet haemocyanin.
    (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

Images

  • ab37305 stained in HeLa cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab37305 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • 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
    All lanes : 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

This product has been referenced in:

  • Salazar VS  et al. Reactivation of a developmental Bmp2 signaling center is required for therapeutic control of the murine periosteal niche. Elife 8:N/A (2019). Read more (PubMed: 30735122) »
  • Wang L  et al. Mechanism of Low Expression of miR-30a-5p on Epithelial-Mesenchymal Transition and Metastasis in Ovarian Cancer. DNA Cell Biol 38:341-351 (2019). Read more (PubMed: 30839226) »
See all 13 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

Submitted Oct 30 2014

Question
Answer

Thank you for contacting us.

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.

We do have other tissue types that can be used as control signal. OurHeLa Nuclear Lysate, (ab14655:https://www.abcam.com/HeLa-Human-epithelial-carcinoma-cell-line-Nuclear-Lysate-ab14655.html) gave the next best signal after MCF7 with ab37305. This product is in stock with next day delivery.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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