Epithelial-Mesenchymal Transition (Beta Catenin, Vimentin) Western Blot Cocktail (ab157392)

Overview

  • Product name
    Epithelial-Mesenchymal Transition (Beta Catenin, Vimentin) Western Blot Cocktail
  • Sample type
    Cell Lysate
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    This Epithelial-Mesenchymal Transition (EMT) western blot cocktail is designed to determine the relative abundance of canonical EMT markers. EMT is a process in which a cell transitions from an epithelial-like (attached, polarized) state to a mesenchymal-like (unattached, migratory) state and such transitions occur during development and metastasis. Underlying the morphological cell changes during EMT are transcription factor programs, biochemical cascades and alterations to the cytoskeleton. Beta-Catenin is a transcription factor in the Wnt signaling pathway and is involved in the regulation of cell adhesion; beta-Catenin is typically more abundant in epithelial-like cells. Vimentin is an intermediate filament typically found in non-epithelial and mesenchymal cells. Alpha Smooth Muscle Actin and GAPDH are also included in this cocktail as loading controls (Alpha Smooth Muscle Actin is reported to be more highly expressed in mesenchymal-like cells, but we find it to be widely expressed in transformed cell lines).

    These four readouts are easily resolved by western blot given their different molecular weights. Because this western blot cocktail is a mix of rabbit and mouse monoclonal primary antibodies.

    A secondary antibody cocktail of HRP conjugated antibodies (anti-mouse and anti-rabbit) is provided.

    Expected and observed MWs:

    • Beta-Catenin 92kDa
    • Vimentin 54kDa
    • Smooth Muscle Actin 42kDa
    • GAPDH 36kDa

     

  • Tested applications
    Suitable for: WBmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab157392 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 at an assay dependent concentration.

Expected and observed MWs:

Beta-Catenin 92kDa

Vimentin 54kDa

Smooth Muscle Actin 42kDa

GAPDH 36kDa

Images

  • Primary antibody: all lanes EMT WB cocktail (ab157392) diluted to 1X in 4% milk/PBS.

    Lane 1. MDA-MB-231 cell lysate
    Lane 2. SH SY5Y cell lysate
    Lane 3. HeLa cell lysate
    Lane 4. Jurkat cell lysate.

    All lysates loaded 20 µg per lane. Lysates were generated in RIPA lysis buffer (ab156034) and briefly boiled in sample buffer before loading on SDS-PAGE.

    Secondary antibody: All lanes using the included HRP Conjugated Secondary Antibody Cocktail (included with ab157392) diluted to 1X in 5% milk/PBS.
  • MCF7 is epithelial-like and MDA-MB-231 cells are mesenchymal-like based on the relative expression of beta-Catenin and Vimentin between the cell lines.

    Primary antibody: all lanes EMT WB cocktail diluted to 1X in 5% milk/PBS.

    Lane 1. MCF7 cell lysate
    Lane 2. MDA-MB-231 cell lysate

    All lysates loaded 20 µg per lane. Lysates were generated in SDS lysis buffer and briefly boiled in sample buffer before loading on SDS-PAGE.

    Secondary antibody: All lanes using the included HRP Conjugated Secondary Antibody Cocktail (included with ab157392) diluted to 1X in 5% milk/PBS.

References

This product has been referenced in:
  • Matsushima H  et al. Sox9 expression in carcinogenesis and its clinical significance in intrahepatic cholangiocarcinoma. Dig Liver Dis 47:1067-75 (2015). WB . Read more (PubMed: 26341967) »

See 1 Publication for this product

Customer reviews and Q&As

Examples of suitable mouse-derived positive controls for product Epithelial-Mesenchymal Transition (Beta Catenin, Vimentin) Western Blot Cocktail (ab157392) include spleen cells, lymphocytes and chondrocytes. For reference, please see:

Al...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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