Overview

  • Product name
    Anti-IRS2 antibody [EP976Y]
    See all IRS2 primary antibodies
  • Description
    Rabbit monoclonal [EP976Y] to IRS2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, Flow Cytmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human IRS2.

  • Positive control
    • 293 cell lysate; human kidney tissue; HeLa; HepG2; Jurkat IF/ICC: SKNSH cell line.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab52606 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 1/5000. Detects a band of approximately 185 kDa (predicted molecular weight: 137 kDa).
ICC/IF 1/100.
Flow Cyt 1/50.
  • Application notes
    Is unsuitable for IHC-P or IP.
  • Target

    • Function
      May mediate the control of various cellular processes by insulin.
    • Sequence similarities
      Contains 1 IRS-type PTB domain.
      Contains 1 PH domain.
    • Post-translational
      modifications
      Phosphorylated upon DNA damage, probably by ATM or ATR.
    • Cellular localization
      Cytoplasm > cytosol.
    • Information by UniProt
    • Database links
    • Alternative names
      • Insulin receptor substrate 2 antibody
      • IRS 2 antibody
      • IRS-2 antibody
      • Irs2 antibody
      • IRS2_HUMAN antibody
      see all

    Images

    • Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling IRS2 with purified ab52606 at 1:50 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling IRS2 with Purified ab52606 at 1:100 dilution (5.9μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Lanes 1-3 : Anti-IRS2 antibody [EP976Y] (ab52606) at 1/1000 dilution (purified)
      Lanes 4-6 : Anti-IRS2 antibody [EP976Y] (ab52606) at 1/5000 dilution (purified)

      Lanes 1 & 4 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lanes 2 & 5 : 293T (Human embryonic kidney epithelial cell) whole cell lysates
      Lanes 3 & 6 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 137 kDa



      Blocking and diluting buffer : 5% NFDM/TBST
    • Unpurified ab52606 stained SKNSH cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52606 at 1/100 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    • Anti-IRS2 antibody [EP976Y] (ab52606) at 1/100000 dilution (unpurified) + 293 cell lysate at 10 µg

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

      Predicted band size: 137 kDa
      Observed band size: 185 kDa
      why is the actual band size different from the predicted?

    • Immunohistochemical staining of paraffin-embedded human kidney using unpurified ab52606 at a dilution of 1/100-1/250.

    References

    This product has been referenced in:
    • Sasaki N  et al. Ganglioside GM1 Contributes to the State of Insulin Resistance in Senescent Human Arterial Endothelial Cells. J Biol Chem 290:25475-86 (2015). WB . Read more (PubMed: 26338710) »
    • El-Ami T  et al. A novel inhibitor of the insulin/IGF signaling pathway protects from age-onset, neurodegeneration-linked proteotoxicity. Aging Cell 13:165-74 (2014). Read more (PubMed: 24261972) »
    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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