Overview

  • Product name

  • Description

    Rabbit polyclonal to ECHS1
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
    Predicted to work with: Chimpanzee, Macaque monkey, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human ECHS1 aa 100-200 conjugated to keyhole limpet haemocyanin.
    Database link: P30084

  • Positive control

    • WB: HepG2 whole cell lysate and human liver, heart and skeletal muscle tissue lysates. ICC/IF: HepG2 cells. IHC-P: Human normal liver tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab136603 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 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 28 kDa (predicted molecular weight: 31 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Relevance

    ECHS1 (Enoyl Coenzyme A hydratase, short chain, 1) catalyzes the hydration of 2-trans-enoyl-coenzyme A (CoA) intermediates to L-3-hydroxyacyl-CoAs, in the second step of the mitochondrial fatty acid beta-oxidation pathway.
  • Cellular localization

    Mitochondrion matrix.
  • Database links

  • Alternative names

    • C80529 antibody
    • EC 4.2.1.17 antibody
    • ECHS 1 antibody
    • Enoyl CoA hydratase 1 antibody
    • Enoyl CoA hydratase mitochondrial antibody
    • Enoyl CoA hydratase short chain 1 mitochondrial antibody
    • Enoyl Coenzyme A hydratase short chain 1 antibody
    • Enoyl Coenzyme A hydratase short chain 1 mitochondrial antibody
    • Mitochondrial short chain enoyl coenzyme A hydratase 1 antibody
    • OTTHUMP00000020811 antibody
    • SCEH antibody
    • Short chain enoyl CoA hydratase antibody
    see all

Images

  • IHC image of ECHS1 staining in Human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab136603, 1µg/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.

  • ICC/IF image of ab136603 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab136603, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) MCF7 cells at 5µg/ml.

  • All lanes : Anti-ECHS1 antibody (ab136603) at 1 µg/ml

    Lane 1 : Liver (Human) Tissue Lysate - adult normal tissue
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : Heart (Human) Tissue Lysate - adult normal tissue
    Lane 4 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 31 kDa
    Observed band size: 28 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 58 kDa (possible non-specific binding)


    Exposure time: 2 minutes


    The band observed at 28 kDa could potentially be a cleaved form of ECHS1 due to the presence of a 27 amino acid transit peptide.

    This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab136603 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

ab136603 has not yet been referenced specifically in any publications.

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