RecombinantRabMAb

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

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22167] to Insulin Receptor beta
  • Suitable for: WB, IHC-P, IP
  • Reacts with: Human

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 & Species

    Application Species
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ELISA
Human
IHC-P
Human
IP
Human
WB
Human
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.
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).

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

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

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

    ab227831 has not yet been referenced specifically in any publications.

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