Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] (ab133561)

Overview

  • Product name

    Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)]
    See all Insulin degrading enzyme / IDE primary antibodies
  • Description

    Rabbit monoclonal [EPR6098(2)] to Insulin degrading enzyme / IDE
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Insulin degrading enzyme/ IDE aa 50-150. The exact sequence is proprietary.
    Database link: P14735

  • Positive control

    • HeLa, HepG2, A375, and K562 cell lysates, Human colon tissue
  • General notes

    A trial size is available to purchase for this antibody.

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133561 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 1/1000 - 1/10000. Detects a band of approximately 120 kDa (predicted molecular weight: 118 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    • Function

      Plays a role in the cellular breakdown of insulin, IAPP, glucagon, bradykinin, kallidin and other peptides, and thereby plays a role in intercellular peptide signaling. Degrades amyloid formed by APP and IAPP. May play a role in the degradation and clearance of naturally secreted amyloid beta-protein by neurons and microglia.
    • Sequence similarities

      Belongs to the peptidase M16 family.
    • Post-translational
      modifications

      The N-terminus is blocked.
    • Cellular localization

      Cytoplasm. Cell surface. Present at the cell surface of neuron cells. The membrane-associated isoform is approximately 5 kDa larger than the known cytosolic isoform.
    • Information by UniProt
    • Database links

    • Alternative names

      • Abeta-degrading protease antibody
      • FLJ35968 antibody
      • Ide antibody
      • IDE_HUMAN antibody
      • Insulin protease antibody
      • Insulin-degrading enzyme antibody
      • Insulinase antibody
      • Insulysin antibody
      • OTTHUMP00000020097 antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: IDE knockout HAP1 cell lysate (20 µg)
      Lane 3: K562 cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab133561 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab133561 was shown to specifically react with IDE in wild-type HAP1 cells. No band was observed when IDE knockout samples were examined. Wild-type and IDE knockout samples were subjected to SDS-PAGE. ab133561 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] (ab133561) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : A375 cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 118 kDa
      Observed band size: 120 kDa
      why is the actual band size different from the predicted?

    • Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Insulin degrading enzyme / IDE with ab133561 antibody at a dilution of 1/50.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Cai H  et al. The Mechanisms of Bushen-Yizhi Formula as a Therapeutic Agent against Alzheimer's Disease. Sci Rep 8:3104 (2018). Read more (PubMed: 29449587) »
    • Huang Y  et al. Chinese Herbal Medicine Xueshuantong Enhances Cerebral Blood Flow and Improves Neural Functions in Alzheimer's Disease Mice. J Alzheimers Dis 63:1089-1107 (2018). Read more (PubMed: 29710701) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    This product is known to not work in this application or species.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Monkey Cell (Kiney)
    Permeabilization
    Yes - 0.1% troton x-100
    Specification
    Kiney
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 15 2016

    Application
    Western blot
    Sample
    Monkey Cell lysate - whole cell (Kidney)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Loading amount
    30 µg
    Specification
    Kidney
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted May 09 2016

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Loading amount
    30 µg
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted May 09 2016

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    Yes - 0.3% Triton x 100: 30 min
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted May 04 2016

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this antibody did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued an aternative free of charge replacement of ab133561 with the order number 1213991.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let me know.

    I wish you the best of luck with your research.

    Read More

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