Insulin R/IGF-I R Heterotetramer [p Tyr1136, p Tyr1163, p Tyr1135, p Tyr1162] Antibody - Cross-reactive

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.

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.

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 Anti­gen Affinity‑purified Polyclonal Antibody (Catalog # AF2507, dark orange filled histogram) or isotype control antibody (Catalog # AB-105-C, blue open histogram), followed by Phy­coerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.

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 (//pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/29367764), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, CyTOF-ready, Flow, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

Insulin R/IGF-I R Heterotetramer [p Tyr1136, p Tyr1163, p Tyr1135, p Tyr1162] Antibody - Cross-reactive Summary

• Description: Antibody detects Insulin R [p Tyr1162, p Tyr1163] / IGF-I R phosphorylated [p Tyr1135, p Tyr1136]
• Immunogen: Phosphopeptide containing human Insulin R Y1162/Y1163 sites
• Modification: p Tyr1136, p Tyr1163, p Tyr1135, p Tyr1162
• Specificity: Detects human Insulin R dually phosphorylated at Y1162 and Y1163, and human IGF-I R dually phosphorylated at Y1135 and Y1136.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunocytochemistry 5-15 ug/mL
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Array

• Insulin R/IGF-I R Heterotetramer

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Insulin R/IGF-I R Heterotetramer Antibody (AF2507)(6)

Publications using AF2507

• Hayashi, SY;Craddock, BP;Miller, WT; Effects of heterologous kinase domains on growth factor receptor specificity Cellular signalling 2024-07-22 [PMID: 39048037] (Immunoprecipitation, Human)
• Pavana Suresh, W. Todd Miller, Erwin London Phospholipid exchange shows insulin receptor activity is supported by both the propensity to form wide bilayers and ordered raft domains Journal of Biological Chemistry 9/1/2021 [PMID: 34324831]
• M Turtoi, M Anghelache, AA Patrascu, C Maxim, I Manduteanu, M Calin, DL Popescu Synthesis, Characterization, and In Vitro Insulin-Mimetic Activity Evaluation of Valine Schiff Base Coordination Compounds of Oxidovanadium(V) Biomedicines, 2021-05-17;9(5):. 2021-05-17 [PMID: 34067862] (Western Blot, Human)
• Benabbou N, Mirshahi P, Bordu C, Faussat A, Tang R, Therwath A, Soria J, Marie J, Mirshahi M A subset of bone marrow stromal cells regulate ATP-binding cassette gene expression via insulin-like growth factor-I in a leukemia cell line. Int J Oncol, 2014-07-29;45(4):1372-80. 2014-07-29 [PMID: 25095896] (ICC, Human)
• Purushothaman , Anurag, Babitz , Stephen, Sanderson , Ralph D Heparanase enhances the insulin receptor signaling pathway to activate extracellular signal-regulated kinase in multiple myeloma. J Biol Chem, 2012-10-09;287(49):41288-96. 2012-10-09 [PMID: 23048032] (Western Blot, Human)
• Iozzo RV, Buraschi S, Genua M Decorin antagonizes IGF receptor I (IGF-IR) function by interfering with IGF-IR activity and attenuating downstream signaling. J. Biol. Chem., 2011-08-12;286(40):34712-21. 2011-08-12 [PMID: 21840990] (Western Blot, Human)