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Anti-Elastin antibody [BA-4] (ab9519)

Key features and details

  • Mouse monoclonal [BA-4] to Elastin
  • Suitable for: WB, IHC-P, IHC-Fr
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-Elastin antibody [BA-4]
    See all Elastin primary antibodies

  • Description

    Mouse monoclonal [BA-4] to Elastin

  • Host species

    Mouse

  • Tested Applications & Species

    Application Species
    IHC-Fr
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    corresponding to Bovine Elastin. Bovine alpha-Elastin.

  • Epitope

    BA-4 is specific for a chemotactically active epitope composed of valine, glycine, alanine, and proline in a molar ratio of approximately 2:2:1:1 (PMID 2429696).

  • Positive control

    • WB: Human skin tissue lysate. IHC-P: FFPE normal Human Duodenum tissue sections IHC-Fr: Human normal skin

  • General notes

     

    Elastin is an important polymeric protein of connective tissue that imparts elasticity to vertebrate elastic tissues.

     

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Elastin antibody [BA-4] (ab9519)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Elastin antibody [BA-4] (ab9519)

    IHC image of Elastin staining in a section of formalin-fixed paraffin-embedded normal human normal duodenum* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab9519, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunohistochemistry (Frozen sections) - Anti-Elastin antibody [BA-4] (ab9519)
    Immunohistochemistry (Frozen sections) - Anti-Elastin antibody [BA-4] (ab9519)

    IHC image of ab9519 staining in 10% formaldehyde fixed frozen tissue section of normal human skin.

    Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with ab9519 (1μg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647)) and DAPI for 1 hour at room temperature.

    The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is derived directly from bound ab9519. 

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Elastin antibody [BA-4] (ab9519)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Elastin antibody [BA-4] (ab9519) This image is courtesy of an anonymous Abreview

    ab9519 staining pig small intestine tissue sections by IHC-P.  Sections were fixed with Bouins and subjected to an enzymatic antigen retreival step.  The primary antibody was diluted 1/200 in a commercially available antibody diluent (replacing the blocking step) and incubated with the sample for 1 hour at 20°C.  A HRP conjugated goat anti-mouse antibody was used as the secondary.

    See Abreview

  • Western blot - Anti-Elastin antibody [BA-4] (ab9519)
    Western blot - Anti-Elastin antibody [BA-4] (ab9519)
    Anti-Elastin antibody [BA-4] (ab9519) at 0.51 µg/ml + Human skin tissue lysate - total protein (ab30166) at 20 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes

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