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

(3 citations)   
  • 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 as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
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.
Data Examples
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) 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 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 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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
Related Research Areas
Citations:

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.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. 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;45(4):1372-80.
  2. 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;287(49):41288-96.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. 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;286(40):34712-21.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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Isotype Controls
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Normal Rabbit IgG Control

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Normal Rabbit IgG Control

Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
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Rabbit IgG HRP-conjugated Antibody

WB HAF008 23  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

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Rabbit IgG APC-conjugated Antibody

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Rabbit IgG PE-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

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Rabbit IgG Fluorescein-conjugated Antibody

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Goat Anti-Rabbit IgG Biotinylated Antibody

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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG Antibody

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Rabbit IgG VisUCyte HRP Polymer Antibody

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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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IGF-I R (Y1135/Y1136) Antibody?

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