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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

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Western blot - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
  • Immunocytochemistry/ Immunofluorescence - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
  • Western blot - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
  • Flow Cytometry (Intracellular) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
  • Immunoprecipitation - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
  • Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15449(B)] to ECH1 - C-terminal
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-ECH1 antibody [EPR15449(B)] - C-terminal
    See all ECH1 primary antibodies

  • Description

    Rabbit monoclonal [EPR15449(B)] to ECH1 - C-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293T, A549, U-2 OS, HeLa and Jurkat cells; Human fetal liver lysate, IHC-P: Human endometrium and kidney tissues. ICC/IF: MCF-7 cells. IP: Human fetal liver lysate. Flow Cyt (intra): Jurkat cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab189255 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
1/10000 - 1/50000. Detects a band of approximately 33 kDa (predicted molecular weight: 36 kDa).
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
IP
1/60.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 33 kDa (predicted molecular weight: 36 kDa).
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
IP
1/60.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Isomerization of 3-trans,5-cis-dienoyl-CoA to 2-trans,4-trans-dienoyl-CoA.

  • Pathway

    Lipid metabolism; fatty acid beta-oxidation.

  • Sequence similarities

    Belongs to the enoyl-CoA hydratase/isomerase family.

  • Cellular localization

    Mitochondrion. Peroxisome.
  • Information by UniProt

  • Database links

    • Alternative names

      • 4)-dienoyl-CoA isomerase antibody
      • 5)-Delta(2 antibody
      • Delta(3 5) Delta(2 4) dienoyl CoA isomerase antibody
      • Delta(3 5) Delta(2 4) dienoyl CoA isomerase mitochondrial antibody
      • Delta(3 antibody
      • Delta3 5 delta2 4 dienoyl CoA isomerase antibody
      • Dienoyl CoA isomerase antibody
      • ECH 1 antibody
      • Ech1 antibody
      • ECH1_HUMAN antibody
      • Enoyl Coenzyme A hydratase 1 antibody
      • Enoyl Coenzyme A hydratase 1 peroxisomal antibody
      • HPXEL antibody
      • mitochondrial antibody
      • MS1034 antibody
      • Peroxisomal enoyl CoA hydratase 1 antibody
      • Peroxisomal enoyl coenzyme A hydratase like protein antibody
      see all

    Images

    • Western blot - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Western blot - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      All lanes : Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : ECH1 knockout HEK293T cell lysate
      Lane 3 : A549 cell lysate
      Lane 4 : U-2 OS cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 36 kDa
      Observed band size: 36 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab189255 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa. 

       ab189255 Anti-ECH1 antibody [EPR15449(B)] was shown to specifically react with ECH1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266748 (knockout cell lysate ab257933) was used.  Wild-type and ECH1 knockout samples were subjected to SDS-PAGE.  ab189255 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively.  Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

      Immunhistochemical analysis of paraffin-embedded human endometrium tissue labeling ECH1 with ab189254 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

      Immunhistochemical analysis of paraffin-embedded human kidney tissue labeling ECH1 with ab189254 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Immunocytochemistry/ Immunofluorescence - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

      Immunfluorescent analysis of 100% methanol(-20℃)-fixed MCF-7 cells labeling ECH1 with ab189254 at 1/250 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 555) secondary antibody at 1/200 dilution. Counter stained with DAPI.

    • Western blot - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Western blot - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      All lanes : Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255) at 1/50000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : A549 cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

      Predicted band size: 36 kDa
      Observed band size: 33 kDa why is the actual band size different from the predicted?

    • Flow Cytometry (Intracellular) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Flow Cytometry (Intracellular) - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

      Intracellular flow cytometric analysis of2% paraformaldehyde-fixed Jurkat cells labeling ECH1 with ab189255 at 1/20 dilution (red) compared to aRabbit monoclonal IgG isotype control (green), followed byGoat anti rabbit IgG (FITC) secondary antibody at 1/150 dilution. 

       

       

    • Immunoprecipitation - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Immunoprecipitation - Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

      Western blot analysis of ECH1 in human fetal liver lysate immunoprecipitated using ab189254 at 1/50 dilution.

      Secondary antibody: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution.

    • Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)
      Anti-ECH1 antibody [EPR15449(B)] - C-terminal (ab189255)

    References (0)

    Publishing research using ab189255? Please let us know so that we can cite the reference in this datasheet.

    ab189255 has not yet been referenced specifically in any publications.

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