Human Phospho-Insulin R (Y1162/Y1163)/
IGF-I R (Y1135/Y1136) Antibody
R&D Systems | Catalog # AF2507
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Immunogen
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Scientific Data Images
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.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.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.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.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.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.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.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.Applications for Human Phospho-Insulin R (Y1162/Y1163)/IGF-I R (Y1135/Y1136) Antibody
CyTOF-ready
Immunocytochemistry
Sample: Immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Intracellular Staining by Flow Cytometry
Sample: HeLa human cervical epithelial carcinoma cell line treated with pervanadate, fixed with paraformaldehyde, and permeabilized with methanol
Western Blot
Sample: Pervanadate-treated A431 human epithelial carcinoma cell line
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Formulation, Preparation, and Storage
Purification
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Shipping
Stability & Storage
- 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.
Calculators
Background: Insulin R/IGF-I R Heterotetramer
Long Name
Additional Insulin R/IGF-I R Heterotetramer Products
Product Documents for Human Phospho-Insulin R (Y1162/Y1163)/IGF-I R (Y1135/Y1136) Antibody
Certificate of Analysis
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Product Specific Notices for Human Phospho-Insulin R (Y1162/Y1163)/IGF-I R (Y1135/Y1136) Antibody
For research use only
Citations for Human Phospho-Insulin R (Y1162/Y1163)/
IGF-I R (Y1135/Y1136) Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Liperfluo
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars