Overview

  • Product name
    Anti-Endothelin 1 antibody
    See all Endothelin 1 primary antibodies
  • Description
    Rabbit polyclonal to Endothelin 1
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Chimpanzee, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human Endothelin 1.

  • Positive control
    • This antibody gave a positive signal in Human Lung and Placenta tissue lysates. It also gave a positive signal in FFPE human placenta tissue sections and JEG3 cell line in IF.

Properties

Applications

Our Abpromise guarantee covers the use of ab113697 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/250. Detects a band of approximately 15 kDa (predicted molecular weight: 24 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

Anti-Endothelin 1 antibody images

  • All lanes : Anti-Endothelin 1 antibody (ab113697) at 1/250 dilution

    Lane 1 : Lung (Human) Tissue Lysate
    Lane 2 : Human placenta tissue lysate - total protein (ab29745)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 24 kDa
    Observed band size : 15 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 80 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutesThe band observed at 15 kDa could potentially be a cleaved form of Endothelin 1 due to the presence of both a signal and pro-peptide.
  • ab113697 stained in JEG3 cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab113697 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature

  • IHC image of Endothelin 1 staining in human placenta formalin fixed paraffin embedded tissue section, 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 20 mins. The section was then incubated with ab113697, 5µg/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.

    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.

References for Anti-Endothelin 1 antibody (ab113697)

This product has been referenced in:
  • Calhan OY & Seyrantepe V Mice with Catalytically Inactive Cathepsin A Display Neurobehavioral Alterations. Behav Neurol 2017:4261873 (2017). IHC-Fr ; Mouse . Read more (PubMed: 28133419) »
  • Daehn I  et al. Endothelial mitochondrial oxidative stress determines podocyte depletion in segmental glomerulosclerosis. J Clin Invest 124:1608-21 (2014). IHC-P . Read more (PubMed: 24590287) »

See all 2 Publications for this product

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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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