Overview

  • Product name
    Anti-Insulin Receptor antibody
    See all Insulin Receptor primary antibodies
  • Description
    Rabbit polyclonal to Insulin Receptor
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Insulin Receptor aa 21-52 (N terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    LLLGAAGHLYPGEVCPGMDIRNNLTRLHELEN


    Database link: P06213
    (Peptide available as ab197200)

  • Positive control
    • WB: SKBR-3 cell lysate and rat and mouse liver tissue lysates. IHC-P: Human breast carcinoma tissue.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Ammonium Sulphate Precipitation
  • Purification notes
    This antibody is prepared by Saturated Ammonium Sulfate (SAS) precipitation followed by dialysis against PBS.
  • Primary antibody notes
    Protein kinases are enzymes that transfer a phosphate group from a phosphate donor, generally the g phosphate of ATP, onto an acceptor amino acid in a substrate protein. By this basic mechanism, protein kinases mediate most of the signal transduction in eukaryotic cells, regulating cellular metabolism, transcription, cell cycle progression, cytoskeletal rearrangement and cell movement, apoptosis, and differentiation. With more than 500 gene products, the protein kinase family is one of the largest families of proteins in eukaryotes. The family has been classified in 8 major groups based on sequence comparison of their tyrosine (PTK) or serine/threonine (STK) kinase catalytic domains. The tyrosine kinase (TK) group is mainly involved in the regulation of cell-cell interactions such as differentiation, adhesion, motility and death. There are currently about 90 TK genes sequenced, 58 are of receptor protein TK (e.g. EGFR, EPH, FGFR, PDGFR, TRK, and VEGFR families), and 32 of cytosolic TK (e.g. ABL, FAK, JAK, and SRC families).
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5500 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
IHC-P 1/50 - 1/100.
WB 1/1000. Detects a band of approximately 165 kDa (predicted molecular weight: 156 kDa).
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    Receptor tyrosine kinase which mediates the pleiotropic actions of insulin. Binding of insulin leads to phosphorylation of several intracellular substrates, including, insulin receptor substrates (IRS1, 2, 3, 4), SHC, GAB1, CBL and other signaling intermediates. Each of these phosphorylated proteins serve as docking proteins for other signaling proteins that contain Src-homology-2 domains (SH2 domain) that specifically recognize different phosphotyrosines residues, including the p85 regulatory subunit of PI3K and SHP2. Phosphorylation of IRSs proteins lead to the activation of two main signaling pathways: the PI3K-AKT/PKB pathway, which is responsible for most of the metabolic actions of insulin, and the Ras-MAPK pathway, which regulates expression of some genes and cooperates with the PI3K pathway to control cell growth and differentiation. Binding of the SH2 domains of PI3K to phosphotyrosines on IRS1 leads to the activation of PI3K and the generation of phosphatidylinositol-(3, 4, 5)-triphosphate (PIP3), a lipid second messenger, which activates several PIP3-dependent serine/threonine kinases, such as PDPK1 and subsequently AKT/PKB. The net effect of this pathway is to produce a translocation of the glucose transporter SLC2A4/GLUT4 from cytoplasmic vesicles to the cell membrane to facilitate glucose transport. Moreover, upon insulin stimulation, activated AKT/PKB is responsible for: anti-apoptotic effect of insulin by inducing phosphorylation of BAD; regulates the expression of gluconeogenic and lipogenic enzymes by controlling the activity of the winged helix or forkhead (FOX) class of transcription factors. Another pathway regulated by PI3K-AKT/PKB activation is mTORC1 signaling pathway which regulates cell growth and metabolism and integrates signals from insulin. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 thereby activating mTORC1 pathway. The Ras/RAF/MAP2K/MAPK pathway is mainly involved in mediating cell growth, survival and cellular differentiation of insulin. Phosphorylated IRS1 recruits GRB2/SOS complex, which triggers the activation of the Ras/RAF/MAP2K/MAPK pathway. In addition to binding insulin, the insulin receptor can bind insulin-like growth factors (IGFI and IGFII). Isoform Short has a higher affinity for IGFII binding. When present in a hybrid receptor with IGF1R, binds IGF1. PubMed:12138094 shows that hybrid receptors composed of IGF1R and INSR isoform Long are activated with a high affinity by IGF1, with low affinity by IGF2 and not significantly activated by insulin, and that hybrid receptors composed of IGF1R and INSR isoform Short are activated by IGF1, IGF2 and insulin. In contrast, PubMed:16831875 shows that hybrid receptors composed of IGF1R and INSR isoform Long and hybrid receptors composed of IGF1R and INSR isoform Short have similar binding characteristics, both bind IGF1 and have a low affinity for insulin.
  • Tissue specificity
    Isoform Long and isoform Short are predominantly expressed in tissue targets of insulin metabolic effects: liver, adipose tissue and skeletal muscle but are also expressed in the peripheral nerve, kidney, pulmonary alveoli, pancreatic acini, placenta vascular endothelium, fibroblasts, monocytes, granulocytes, erythrocytes and skin. Isoform Short is preferentially expressed in fetal cells such as fetal fibroblasts, muscle, liver and kidney. Found as a hybrid receptor with IGF1R in muscle, heart, kidney, adipose tissue, skeletal muscle, hepatoma, fibroblasts, spleen and placenta (at protein level). Overexpressed in several tumors, including breast, colon, lung, ovary, and thyroid carcinomas.
  • Involvement in disease
    Rabson-Mendenhall syndrome
    Leprechaunism
    Diabetes mellitus, non-insulin-dependent
    Familial hyperinsulinemic hypoglycemia 5
    Insulin-resistant diabetes mellitus with acanthosis nigricans type A
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family. Insulin receptor subfamily.
    Contains 3 fibronectin type-III domains.
    Contains 1 protein kinase domain.
  • Domain
    The tetrameric insulin receptor binds insulin via non-identical regions from two alpha chains, primarily via the C-terminal region of the first INSR alpha chain. Residues from the leucine-rich N-terminus of the other INSR alpha chain also contribute to this insulin binding site. A secondary insulin-binding site is formed by residues at the junction of fibronectin type-III domain 1 and 2.
  • Post-translational
    modifications
    After being transported from the endoplasmic reticulum to the Golgi apparatus, the single glycosylated precursor is further glycosylated and then cleaved, followed by its transport to the plasma membrane.
    Autophosphorylated on tyrosine residues in response to insulin. Phosphorylation of Tyr-999 is required for binding to IRS1, SHC1 and STAT5B. Dephosphorylated by PTPRE at Tyr-999, Tyr-1185, Tyr-1189 and Tyr-1190. Dephosphorylated by PTPRF and PTPN1. Dephosphorylated by PTPN2; down-regulates insulin-induced signaling.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD220 antibody
    • HHF5 antibody
    • human insulin receptor antibody
    • Insr antibody
    • INSR_HUMAN antibody
    • Insulin receptor subunit beta antibody
    • IR 1 antibody
    • IR antibody
    • IR-1 antibody
    • IR1 antibody
    see all

Anti-Insulin Receptor antibody images

  • Anti-Insulin Receptor antibody (ab5500) at 1/1000 dilution + SKBR-3 cell lysate at 35 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution

    Predicted band size : 156 kDa

    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Insulin Receptor using ab5500. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 38°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hour at 37°C. A peroxidase-conjugated goat anti-rabbit polyclonal (ready to use) was used as the secondary antibody.

  • ab5500 staining Insulin Receptor in Human WBCs by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 2% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/250 in PBS + 2% BSA) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (ab150077) (1/500) was used as the secondary antibody.

    See Abreview

  • Anti-Insulin Receptor antibody (ab5500) at 1/2000 dilution + Mouse liver tissue lysate at 20 µg

    Secondary
    Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size : 156 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-Insulin Receptor antibody (ab5500) at 1/2000 dilution + Rat liver tissue lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size : 156 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

References for Anti-Insulin Receptor antibody (ab5500)

This product has been referenced in:
  • Li L  et al. Absence of renal enlargement in fructose-fed proximal-tubule-select insulin receptor (IR), insulin-like-growth factor receptor (IGF1R) double knockout mice. Physiol Rep 4:N/A (2016). Mouse . Read more (PubMed: 27923977) »
  • Krüger J  et al. Targeting density-enhanced phosphatase-1 (DEP-1) with antisense oligonucleotides improves the metabolic phenotype in high-fat diet-fed mice. Cell Commun Signal 11:49 (2013). Read more (PubMed: 23889985) »

See all 2 Publications for this product

Product Wall

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (pancreas)
Antigen retrieval step
None - Buffer/Enzyme Used: HistoVTONE(nakaitesque)
Permeabilization
Yes - tween20
Specification
pancreas
Blocking step
Blocking One (nakalaitesque cat:03953-6) as blocking agent for 2 hour(s) and 0 minute(s) · Temperature: 25°C
Fixative
Formaldehyde
Username

takai shingo

Verified customer

Submitted Jul 18 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (Kidney tissue)
Gel Running Conditions
Reduced Denaturing (4-20% gel (Bio-Rad))
Loading amount
25 µg
Specification
Kidney tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 15 2015

Thank you for contacting us.

The immunogen sequence homology of this antibody with IGF-1 receptor is 61% so I don't think the antibody would cross react with IGF1 receptor.

I hope this information is helpful to you. Please do not hesi...

Read More

Thank you for this information. We have not tested this product in rat, atlhough it is predicted to work based on homology with the immunogen it is not guaranteed. You may improve the result by increasing your primary antibody dilution to 1/10-1/50 and...

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Bone marrow)
Specification
Bone marrow
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (WBCs)
Specification
WBCs
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 12 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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