Anti-Insulin Receptor antibody (ab137747)
Key features and details
- Rabbit polyclonal to Insulin Receptor
- Suitable for: WB, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-Insulin Receptor antibody
See all Insulin Receptor primary antibodies -
Description
Rabbit polyclonal to Insulin Receptor -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment, corresponding to a region within internal sequence amino acids 939-1347 of Human Insulin Receptor
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Positive control
- WB: Wild-type HepG2, HeLa, HEK-293T, NIH-3T3 and PC-12 cell lysates. IHC-P: Human gastric cancer and mouse prostate tissues.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.00
Preservative: 0.025% Proclin 300
Constituents: 79% PBS, 20% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab137747 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 |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 156 kDa.
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IHC-P |
Use at an assay dependent concentration.
Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min. |
Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 156 kDa. |
IHC-P
Use at an assay dependent concentration. Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min. |
Target
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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
Leprechaunism
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
modificationsAfter 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
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Database links
- Entrez Gene: 3643 Human
- Entrez Gene: 16337 Mouse
- Entrez Gene: 24954 Rat
- Omim: 147670 Human
- SwissProt: P06213 Human
- SwissProt: P15208 Mouse
- SwissProt: P15127 Rat
- Unigene: 465744 Human
see all -
Alternative names
- CD220 antibody
- HHF5 antibody
- human insulin receptor antibody
see all
Images
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All lanes : Anti-Insulin Receptor antibody (ab137747) at 1/1000 dilution
Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : INSR knockout HepG2 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : HDLM-2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 156 kDa
Observed band size: 200,85 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Insulin Receptor antibody staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab137747 was shown to bind specifically to Insulin Receptor. A band was observed at 85/200 kDa in wild-type HepG2 cell lysates with no signal observed at this size in INSR knockout cell line ab282827 (knockout cell lysate ab283051). Pro-form of INSR is observed at 200 kDa and the mature form observed at 90 kDa. To generate this image, wild-type and INSR knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labelling INSR protein at cytosol with ab137747 at 1/500 dilution
Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min.
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Anti-Insulin Receptor antibody (ab137747) at 1/1000 dilution + HEK-293T at 30 µg
Secondary
HRP-conjugated anti-rabbit IgG antibody
Predicted band size: 156 kDa5% SDS PAGE
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Anti-Insulin Receptor antibody (ab137747) at 1/10000 dilution + HeLa whole cell lysate at 30 µg
Predicted band size: 156 kDa
5% SDS PAGE -
Immunohistochemical analysis of paraffin-embedded mouse prostate tissue labelling Insulin receptor protein at membrane with ab137747 at 1/500 dilution.
Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min. -
Anti-Insulin Receptor antibody (ab137747) at 1/1000 dilution + NIH-3T3 at 30 µg
Secondary
HRP-conjugated anti-rabbit IgG antibody
Predicted band size: 156 kDa5% SDS PAGE
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All lanes : Anti-Insulin Receptor antibody (ab137747) at 1/1000 dilution
Lane 1 : PC-12
Lane 2 : Rat2
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody
Predicted band size: 156 kDa5% SDS PAGE
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab137747 has been referenced in 7 publications.
- Jiang A et al. Free Fatty Acid Impairs Myogenic Differentiation through the AMPKα-MicroRNA 206 Pathway. Mol Cell Biol 42:e0032721 (2022). PubMed: 34694913
- Han L et al. Mechanism exploration of Gouqi-wentang formula against type 2 diabetes mellitus by phytochemistry and network pharmacology-based analysis and biological validation. Chin Med 16:93 (2021). PubMed: 34579756
- Yun C et al. Pilose antler peptide promotes osteoblast proliferation, differentiation and mineralization via the insulin signaling pathway. Exp Ther Med 19:923-930 (2020). PubMed: 32010253
- Wang L et al. A MicroRNA Linking Human Positive Selection and Metabolic Disorders. Cell 183:684-701.e14 (2020). PubMed: 33058756
- Yang Y et al. Insulin receptor is implicated in triple-negative breast cancer by decreasing cell mobility. J Biomed Res 35:189-196 (2020). PubMed: 33911052
- Ratai O et al. NCS-1 Deficiency Is Associated With Obesity and Diabetes Type 2 in Mice. Front Mol Neurosci 12:78 (2019). PubMed: 31001084
- Jackson R et al. Paracrine Engineering of Human Cardiac Stem Cells With Insulin-Like Growth Factor 1 Enhances Myocardial Repair. J Am Heart Assoc 4:e002104 (2015). WB, IHC ; Mouse, Human . PubMed: 26363004