Home / Insulin R/IGF-I R Heterotetramer / Human Phospho-Insulin R (Y1162/Y1163)/
IGF-I R (Y1135/Y1136) Antibody

Human Phospho-Insulin R (Y1162/Y1163)/
IGF-I R (Y1135/Y1136) Antibody

Catalog #: AF2507
6 Citations Datasheet
Catalog # Availability Size / Price Qty
AF2507
AF2507-SP
Detection of Human Phospho‑Insulin R (Y1162/Y1163) and Phospho‑IGF‑I R (Y1135/1136) by Western Blot.
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Citations (6)
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Human Phospho-Insulin R (Y1162/Y1163)/
IGF-I R (Y1135/Y1136) Antibody Summary

Species Reactivity
Human
Specificity
Detects human Insulin R dually phosphorylated at Y1162 and Y1163, and human IGF-I R dually phosphorylated at Y1135 and Y1136.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Phosphopeptide containing human Insulin R Y1162/Y1163 sites
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
5-15 µg/mL
See below
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Phospho-Insulin R (Y1162/Y1163) and Phospho-IGF-I R (Y1135/1136) antibody by Western Blot. View Larger

Detection of Human Phospho‑Insulin R (Y1162/Y1163) and Phospho‑IGF‑I R (Y1135/1136) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 µM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-Insulin R (Y1162/Y1163)/IGF-I R (Y1135/Y1136) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2507), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-Insulin R (Y1162/Y1163) at 95 kDa and Phospho-IGF-I R (Y1135/1136) at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry Phospho-Insulin R (Y1162/1163)/IGF-I R (Y1135/1136) antibody in A431 Human Cell Line by Immunocytochemistry (ICC). View Larger

Phospho‑Insulin R (Y1162/1163)/IGF-I R (Y1135/1136) in A431 Human Cell Line. Insulin R phosphorylated at Y1162/1163 and IGF-I R phsophorylated at Y1135/1136 were detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-Insulin R (Y1162/1163)/IGF-I R (Y1135/1136) Cross-reactive Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2507) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Intracellular Staining by Flow Cytometry Detection of Insulin R/CD220 antibody in HeLa Human Cell Line antibody by Flow Cytometry. View Larger

Detection of Insulin R/CD220 in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was unstimulated (light orange open histogram) or treated with 100 µM pervanadate for 15 minutes, then stained with Rabbit Anti-Human Phospho-Insulin R (Y1162/1163)/IGF-I R (Y1135/1136) cross-reactive Anti-gen Affinity-purified Polyclonal Antibody (Catalog # AF2507, dark orange filled histogram) or isotype control antibody (Catalog # AB-105-C, blue open histogram), followed by Phy-coerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.

Immunohistochemistry Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry View Larger

Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry Eighty-seven percent of patients with invasive breast cancer have activated Insulin/IGF1 receptors. a Immunohistochemical staining of invasive breast cancer tissue serial sections stained for phospho-Insulin/IGF1 receptor and CD163. Scale bars, 100 μm and 50 μm. b Upper diagram: Pie diagram representing the percentage of phospho-Insulin/IGF1 receptor positive (red) and negative (green) tumors assessed in tissue microarray (TMA) containing biopsies from 90 consented patients with invasive breast cancer. Lower diagrams: represent the percentage of phospho-Insulin/IGF1 receptor positive (red) and negative (green) tumors of the molecular subsets, TNBC, HR+, and HER2+. c Expression levels of Igf-1, Igf-2, cd163, and mrc1 associated with survival in breast cancer patients Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry View Larger

Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry 75% of breast cancer patients have activated Insulin/IGF1 receptors and Insulin/IGF-1 receptor activation positively correlates with macrophage infiltration and advanced tumor stage. a Serial sections of biopsies from non-malignant breast tissue immunohistochemically stained for phospho-Insulin/IGF1 receptor, CD163 and CD68. Scale bars, 100 μm and 50 μm. b and c Serial sections of biopsies from breast cancer patients immunohistochemically stained for phospho-insulin/IGF1 receptor, CD163, and CD68. Scale bars, 100 μm and 50 μm. d Bar graph depicting the quantification of CD68 and CD163 positive macrophages in non-malignant breast tissue and breast cancer tissue samples. Error bars represent s.d. (n = 3); * two-tailed p-value ≤ 0.05, *** two-tailed p-value ≤ 0.005 using a student’s t-test. e Pie diagram representing the percentage of phospho-Insulin/IGF-1 receptor positive (red) and negative (green) tumors assessed in a tissue microarray containing biopsies from 75 breast cancer patients. f Contingency table and results from statistical analysis showing a positive correlation between phospho-Insulin/IGF-1 receptor expression in breast tumors and increased CD163+ macrophage infiltration. Chi-square = 4.37; p = 0.04. g Contingency table and results from statistical analysis showing a positive correlation between phospho-Insulin/IGF1 receptor and CD163+ macrophages co-expression and tumor stage. Chi-square = 4.89; p = 0.03 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry View Larger

Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry 75% of breast cancer patients have activated Insulin/IGF1 receptors and Insulin/IGF-1 receptor activation positively correlates with macrophage infiltration and advanced tumor stage. a Serial sections of biopsies from non-malignant breast tissue immunohistochemically stained for phospho-Insulin/IGF1 receptor, CD163 and CD68. Scale bars, 100 μm and 50 μm. b and c Serial sections of biopsies from breast cancer patients immunohistochemically stained for phospho-insulin/IGF1 receptor, CD163, and CD68. Scale bars, 100 μm and 50 μm. d Bar graph depicting the quantification of CD68 and CD163 positive macrophages in non-malignant breast tissue and breast cancer tissue samples. Error bars represent s.d. (n = 3); * two-tailed p-value ≤ 0.05, *** two-tailed p-value ≤ 0.005 using a student’s t-test. e Pie diagram representing the percentage of phospho-Insulin/IGF-1 receptor positive (red) and negative (green) tumors assessed in a tissue microarray containing biopsies from 75 breast cancer patients. f Contingency table and results from statistical analysis showing a positive correlation between phospho-Insulin/IGF-1 receptor expression in breast tumors and increased CD163+ macrophage infiltration. Chi-square = 4.37; p = 0.04. g Contingency table and results from statistical analysis showing a positive correlation between phospho-Insulin/IGF1 receptor and CD163+ macrophages co-expression and tumor stage. Chi-square = 4.89; p = 0.03 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry View Larger

Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry Combined treatment of IGF blocking antibody with paclitaxel decreases breast cancer proliferation and metastasis in Py230 model. a Py230 luciferase cells were orthotopically implanted into the third mammary fatpad of syngeneic C57BL/6 recipient mice and mice were treated, starting when tumors reached between 5–8 mm2, twice a week i.p., with control IgG antibody, IGF blocking antibody xentuzumab (100 mg/kg), paclitaxel (100 mg/kg), or a combination of xentuzumab with paclitaxel (n = 8 mice per group). b Immunohistochemical staining of phospho-insulin/IGF-1R in breast tumors treated with IgG (control), paclitaxel, xentuzumab or paclitaxel + xentuzumab. Scale bars 100 μm and 50 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry View Larger

Detection of Human Human Phospho-Insulin R (Y1162/Y1163)/ IGF-I R (Y1135/Y1136) Antibody by Immunohistochemistry 75% of breast cancer patients have activated Insulin/IGF1 receptors and Insulin/IGF-1 receptor activation positively correlates with macrophage infiltration and advanced tumor stage. a Serial sections of biopsies from non-malignant breast tissue immunohistochemically stained for phospho-Insulin/IGF1 receptor, CD163 and CD68. Scale bars, 100 μm and 50 μm. b and c Serial sections of biopsies from breast cancer patients immunohistochemically stained for phospho-insulin/IGF1 receptor, CD163, and CD68. Scale bars, 100 μm and 50 μm. d Bar graph depicting the quantification of CD68 and CD163 positive macrophages in non-malignant breast tissue and breast cancer tissue samples. Error bars represent s.d. (n = 3); * two-tailed p-value ≤ 0.05, *** two-tailed p-value ≤ 0.005 using a student’s t-test. e Pie diagram representing the percentage of phospho-Insulin/IGF-1 receptor positive (red) and negative (green) tumors assessed in a tissue microarray containing biopsies from 75 breast cancer patients. f Contingency table and results from statistical analysis showing a positive correlation between phospho-Insulin/IGF-1 receptor expression in breast tumors and increased CD163+ macrophage infiltration. Chi-square = 4.37; p = 0.04. g Contingency table and results from statistical analysis showing a positive correlation between phospho-Insulin/IGF1 receptor and CD163+ macrophages co-expression and tumor stage. Chi-square = 4.89; p = 0.03 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Insulin R/IGF-I R Heterotetramer

The heterotetrameric receptors for insulin (INS R) and IGF-I (IGF-I R) are receptor tyrosine kinases that consist of two ligand-binding alpha subunits and two beta subunits. Ligand binding induces autophosphorylation on multiple tyrosine residues of beta  subunits. Phosphorylation of Tyr1162 and 1163 on INS R and Tyr1135 and 1136 on IGF‑I R stimulates intrinsic kinase activity.

Long Name
Insulin Receptor
Alternate Names
Insulin R/IGF-I R Heterotetramer

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Citations for Human Phospho-Insulin R (Y1162/Y1163)/
IGF-I R (Y1135/Y1136) Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. Phospholipid exchange shows insulin receptor activity is supported by both the propensity to form wide bilayers and ordered raft domains
    Authors: Pavana Suresh, W. Todd Miller, Erwin London
    Journal of Biological Chemistry
  2. Effects of heterologous kinase domains on growth factor receptor specificity
    Authors: Hayashi, SY;Craddock, BP;Miller, WT;
    Cellular signalling
    Species: Human
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  3. Synthesis, Characterization, and In Vitro Insulin-Mimetic Activity Evaluation of Valine Schiff Base Coordination Compounds of Oxidovanadium(V)
    Authors: M Turtoi, M Anghelache, AA Patrascu, C Maxim, I Manduteanu, M Calin, DL Popescu
    Biomedicines, 2021-05-17;9(5):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. A subset of bone marrow stromal cells regulate ATP-binding cassette gene expression via insulin-like growth factor-I in a leukemia cell line.
    Authors: Benabbou N, Mirshahi P, Bordu C, Faussat A, Tang R, Therwath A, Soria J, Marie J, Mirshahi M
    Int J Oncol, 2014-07-29;45(4):1372-80.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  5. Heparanase enhances the insulin receptor signaling pathway to activate extracellular signal-regulated kinase in multiple myeloma.
    Authors: Purushothaman, Anurag, Babitz, Stephen, Sanderson, Ralph D
    J Biol Chem, 2012-10-09;287(49):41288-96.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. Decorin antagonizes IGF receptor I (IGF-IR) function by interfering with IGF-IR activity and attenuating downstream signaling.
    Authors: Iozzo RV, Buraschi S, Genua M
    J. Biol. Chem., 2011-08-12;286(40):34712-21.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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IGF-I R (Y1135/Y1136) Antibody

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