Recombinant
RabMAb

Recombinant Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)

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Overview

  • Product name

    Anti-Insulin Receptor beta antibody [EPR22167]
    See all Insulin Receptor beta primary antibodies

  • Description

    Rabbit monoclonal [EPR22167] to Insulin Receptor beta

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Insulin Receptor beta aa 950 to the C-terminus. The exact sequence is proprietary.
    Database link: P06213

  • Positive control

    • WB: HepG2, HEK-293T, LNCaP, HeLa and MCF7 whole cell lysates; Human fetal brain and liver lysates. IHC-P: Human kidney and kidney carcinoma tissues. IP: HepG2 whole cell lysate.

  • 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

Our Abpromise guarantee covers the use of ab227831 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. Detects a band of approximately 210, 95, 49 kDa (predicted molecular weight: 156 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.

Target

  • Relevance

    Insulin receptor mediates the biological activities of insulin by regulating multiple signaling pathways through activation of a series of phosphorylation cascades. The human insulin receptor is a heterotetrameric membrane glycoprotein consisting of disulfide-linked subunits in a ß-a-a-ß configuration. The ß-subunit (95kDa) possesses a single transmembrane domain with tyrosine kinase acivity, whereas the a-subunit (135kDa) is completely extracellular. The alpha subunits each contain insulin binding sites and are entirely extracellular in localization. The beta subunits each possess an extracellular domain, a single transmembrane domain, and a cytoplasmic tyrosine kinase domain. Binding of insulin to the alpha subunits induces a conformation change in the receptor which activates the kinase domain, stimulating tyrosine autophosphorylation of the receptor and tyrosine phosphorylation of at least five different insulin receptor substrates designated IRS-1-4, and Shc.

  • Cellular localization

    Membrane; Single pass type I membrane protein.

  • Database links

    • Alternative names

      • CD220 antibody
      • HHF5 antibody
      • HIR B antibody
      • INSR antibody
      • Insulin receptor antibody
      • Insulin receptor subunit beta antibody
      • IR antibody
      see all

    Images

    • Western blot - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)
      Western blot - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)
      All lanes : Anti-Insulin Receptor beta antibody [EPR22167] (ab227831) at 1/1000 dilution

      Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
      Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
      Lane 3 : LNCaP (human prostate cancer cell line) whole cell lysate at 10 µg
      Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
      Lane 5 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 10 µg
      Lane 6 : Human fetal brain lysate at 10 µg
      Lane 7 : Human liver lysate at 10 µg

      Secondary
      Lanes 1-5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lanes 6-7 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 156 kDa



      Exposure time : Lanes 1-2: 48 seconds; Lanes 3: 59 seconds; Lanes 4&6-7: 70 seconds; Lane 5: 37 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      The 210 kDa band is the pro-insulin receptor, while the 95 kDa band is the insulin receptor beta subunit (PMID: 28765322, PMID: 28915606).

      The 45-68 kDa bands are proteolytic cleavage fragments (PMID: 28915606, PMID: 6693383, PMID: 6315728).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)

      Immunohistochemical analysis of paraffin-embedded human kidney (Panel A) and kidney carcinoma (Panel B) tissues labeling Insulin Receptor beta with ab227831 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in human kidney tubules (panel A). Positive staining in endothelium of blood vessels in human kidney carcinoma (panel B), PMID: 25864925, PMID: 20182859. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)

      Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling Insulin Receptor beta with ab227831 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in endothelium of blood vessels in human glioma (PMID: 26136493) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)
      Immunoprecipitation - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)

      Insulin Receptor beta was immunoprecipitated from 0.35 mg of HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate with ab227831 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227831 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: HepG2 whole cell lysate 10 µg (Input). 

      Lane 2: 227831 IP in HepG2 whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of 227831 in HepG2 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 5 seconds.

      The 210 kDa band is the pro-Insulin receptor, while the 95 kDa band is the insulin receptor beta subunit (PMID: 28765322, PMID: 28915606).

      The 45-68 kDa bands are proteolytic cleavage fragments (PMID: 28915606, PMID: 6693383, PMID: 6315728).

    • Immunoprecipitation - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)
      Immunoprecipitation - Anti-Insulin Receptor beta antibody [EPR22167] (ab227831)

      Insulin Receptor beta was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with ab227831 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227831 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: HEK-293T whole cell lysate 10 µg (Input). 

      Lane 2: 227831 IP in HEK-293T whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of 227831 in HEK-293T whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 5 seconds.

      The 210 kDa band is the pro-Insulin receptor, while the 95 kDa band is the insulin receptor beta subunit (PMID: 28765322, PMID: 28915606).

      The 45-68 kDa bands are proteolytic cleavage fragments (PMID: 28915606, PMID: 6693383, PMID: 6315728).

    References

    ab227831 has not yet been referenced specifically in any publications.

    Publishing research using ab227831? Please let us know so that we can cite the reference in this datasheet.

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