Anti-Insulin Receptor (phospho Y1361) antibody (ab60946)

Rabbit polyclonal Insulin Receptor (phospho Y1361) antibody. Validated in WB, ELISA, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 9 publication(s). Independently reviewed in 3 review(s).


  • Product name

    Anti-Insulin Receptor (phospho Y1361) antibody
    See all Insulin Receptor primary antibodies
  • Description

    Rabbit polyclonal to Insulin Receptor (phospho Y1361)
  • Host species

  • Specificity

    ab60946 detects endogenous levels of Insulin Receptor only when phosphorylated at tyrosine 1361. In mouse the modification site is tyrosine 1351 and in rat it is tyrosine 1362.

  • Tested applications

    Suitable for: IHC-P, ELISA, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Insulin Receptor. The immunogen is a synthesized peptide derived from human Insulin Receptor around the phosphorylation site of Tyr1361.
    Database link: P06213

  • Positive control

    • WB: CHO treated with 0.01 U/mL Insulin for 15 minutes lysate, 293 treated with 0.01 U/mL Insulin for 15 minutes lysate. ICC/IF: Human white blood cells. IHC-P: Human spleen, rat brain.



Our Abpromise guarantee covers the use of ab60946 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 Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ELISA 1/10000.
WB 1/500 - 1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 156 kDa).
ICC/IF 1/500.


  • 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
    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

    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


  • All lanes : Anti-Insulin Receptor (phospho Y1361) antibody (ab60946) at 1/1000 dilution

    Lane 1 : CHO untreated lysate
    Lane 2 : CHO treated with 0.01 U/mL Insulin for 15 minutes lysate
    Lane 3 : 293 untreated lysate
    Lane 4 : 293 treated with 0.01 U/mL Insulin for 15 minutes lysate

    Lysates/proteins at 20 µg per lane.

    All lanes : Goat Anti-Rabbit IgG (H+L) HRP at 1/10000 dilution

    Predicted band size: 156 kDa

    Loading control: Beta Actin

  • ab60946 staining the insulin receptor in Human white blood cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with Triton X-100 (0.1%) and blocked with 2% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/500 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

  • ab60946 (1:10000) antibody detects endogenous levels of Insulin Receptor only when phosphorylated at Tyr1361.

  • Ab60946 staining human spleen. Staining is localized to the membrane, with some light staining in the cytoplasm.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, citrate pH 6.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be requ
  • ab60946 staining Insulin Receptor in Rat brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 8% Milk for 30 minutes at 37°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in diluent) for 18 hours at 4°C. A detection reagent was used to detect antibody staining.

    See Abreview

  • All lanes : Anti-Insulin Receptor (phospho Y1361) antibody (ab60946) at 1/500 dilution

    Lane 1 : 293 cell extracts treated
    with Heat shock
    Lane 2 : 293 cell extracts treated
    with Heat shock with immunising peptide

    Predicted band size: 156 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?


This product has been referenced in:

  • Li H  et al. Dioscin attenuates high-fat diet-induced insulin resistance of adipose tissue through the IRS-1/PI3K/Akt signaling pathway. Mol Med Rep 19:1230-1237 (2019). Read more (PubMed: 30483735) »
  • Guo HH  et al. Liver-target nanotechnology facilitates berberine to ameliorate cardio-metabolic diseases. Nat Commun 10:1981 (2019). Read more (PubMed: 31040273) »
See all 12 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Immunohistochemistry (Frozen sections)
Mouse Tissue sections (5um Aorta Cryosections)
Yes - 0.1% Triton-X 100 for 20min
5um Aorta Cryosections
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 22°C

Victoria Tokarz

Verified customer

Submitted Jul 14 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Rat Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate-Na, 0.05%Tween20, pH6.0., and cycle was on 120C 10 min using autoclave
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 8% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Jun 01 2012


Thank you for submitting an Abreview for Anti-Insulin Receptor (phospho Y1361) antibody (ab60946).

We value all of our Abreviews, whether positive or negative, as they are useful resources for other researchers. You seem to have achieved some very nice staining with this antibody.In order to publish this Abreview, there are a few more pieces of information we need.Could you please let me know which buffer was used when performing the antigen retrieval using the autoclave and what cycle was used.

If you could let me know and I will update the Abreview and publish it. Please let me know if there are any questions. Thank you again!

Read More
Immunocytochemistry/ Immunofluorescence
Human Cell (WBCs)
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 11 2012

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