Recombinant Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)
![Western blot - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)](https://www.abcam.com/ps/products/263/ab263907/Images/ab263907-513043-anti-igf1-receptor-antibody-epr23027-80-western-blot-wildtype-mcf7-igf1r-knockout-mcf7-hela-hdlm2.jpg "Western blot - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23027-80] to IGF1 Receptor
- Suitable for: WB, IP, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IGF1 Receptor antibody [EPR23027-80]
See all IGF1 Receptor primary antibodies -
Description
Rabbit monoclonal [EPR23027-80] to IGF1 Receptor -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, Flow Cyt, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A431, HepG2, HeLa and MDA-MB-231 lysates. ICC/IF: MCF7 cells. Flow Cyt: MCF7 cells. IP: MCF7 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23027-80 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab263907 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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 130, 200 kDa (predicted molecular weight: 154 kDa).
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| IP |
1/30.
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| Flow Cyt |
1/500.
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| ICC/IF |
1/100.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 130, 200 kDa (predicted molecular weight: 154 kDa). |
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IP
1/30. |
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Flow Cyt
1/500. |
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ICC/IF
1/100. |
Target
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Function
Receptor tyrosine kinase which mediates actions of insulin-like growth factor 1 (IGF1). Binds IGF1 with high affinity and IGF2 and insulin (INS) with a lower affinity. The activated IGF1R is involved in cell growth and survival control. IGF1R is crucial for tumor transformation and survival of malignant cell. Ligand binding activates the receptor kinase, leading to receptor autophosphorylation, and tyrosines phosphorylation of multiple substrates, that function as signaling adapter proteins including, the insulin-receptor substrates (IRS1/2), Shc and 14-3-3 proteins. Phosphorylation of IRSs proteins lead to the activation of two main signaling pathways: the PI3K-AKT/PKB pathway and the Ras-MAPK pathway. The result of activating the MAPK pathway is increased cellular proliferation, whereas activating the PI3K pathway inhibits apoptosis and stimulates protein synthesis. Phosphorylated IRS1 can activate the 85 kDa regulatory subunit of PI3K (PIK3R1), leading to activation of several downstream substrates, including protein AKT/PKB. AKT phosphorylation, in turn, enhances protein synthesis through mTOR activation and triggers the antiapoptotic effects of IGFIR through phosphorylation and inactivation of BAD. In parallel to PI3K-driven signaling, recruitment of Grb2/SOS by phosphorylated IRS1 or Shc leads to recruitment of Ras and activation of the ras-MAPK pathway. In addition to these two main signaling pathways IGF1R signals also through the Janus kinase/signal transducer and activator of transcription pathway (JAK/STAT). Phosphorylation of JAK proteins can lead to phosphorylation/activation of signal transducers and activators of transcription (STAT) proteins. In particular activation of STAT3, may be essential for the transforming activity of IGF1R. The JAK/STAT pathway activates gene transcription and may be responsible for the transforming activity. JNK kinases can also be activated by the IGF1R. IGF1 exerts inhibiting activities on JNK activation via phosphorylation and inhibition of MAP3K5/ASK1, which is able to directly associate with the IGF1R.
When present in a hybrid receptor with INSR, 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
Found as a hybrid receptor with INSR in muscle, heart, kidney, adipose tissue, skeletal muscle, hepatoma, fibroblasts, spleen and placenta (at protein level). Expressed in a variety of tissues. Overexpressed in tumors, including melanomas, cancers of the colon, pancreas prostate and kidney. -
Involvement in disease
Insulin-like growth factor 1 resistance -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. Insulin receptor subfamily.
Contains 4 fibronectin type-III domains.
Contains 1 protein kinase domain. -
Post-translational
modificationsAutophosphorylated on tyrosine residues in response to ligand binding. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit. Autophosphorylation occurs in a sequential manner; Tyr-1165 is predominantly phosphorylated first, followed by phosphorylation of Tyr-1161 and Tyr-1166. While every single phosphorylation increases kinase activity, all three tyrosine residues in the kinase activation loop (Tyr-1165, Tyr-1161 and Tyr-1166) have to be phosphorylated for optimal activity. Can be autophosphorylated at additional tyrosine residues (in vitro). Autophosphorylated is followed by phosphorylation of juxtamembrane tyrosines and C-terminal serines. Phosphorylation of Tyr-980 is required for IRS1- and SHC1-binding. Phosphorylation of Ser-1278 by GSK-3beta restrains kinase activity and promotes cell surface expression, it requires a priming phosphorylation at Ser-1282. Dephosphorylated by PTPN1.
Polyubiquitinated at Lys-1168 and Lys-1171 through both 'Lys-48' and 'Lys-29' linkages, promoting receptor endocytosis and subsequent degradation by the proteasome. Ubiquitination is facilitated by pre-existing phosphorylation.
Sumoylated with SUMO1.
Controlled by regulated intramembrane proteolysis (RIP). Undergoes metalloprotease-dependent constitutive ectodomain shedding to produce a membrane-anchored 52 kDa C-Terminal fragment which is further processed by presenilin gamma-secretase to yield an intracellular 50 kDa fragment. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 3480 Human
- Omim: 147370 Human
- SwissProt: P08069 Human
- Unigene: 643120 Human
- Unigene: 714012 Human
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Alternative names
- CD221 antibody
- CD221 antigen antibody
- IGF 1 receptor antibody
see all
Images
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Western blot - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)All lanes : Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : IGF1R knockout MCF7 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : HDLM-2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 154 kDa
Observed band size: 105-125 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-IGF1 Receptor antibody [EPR23027-80] 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, ab263907 was shown to bind specifically to IGF1 Receptor. A band was observed at 105-125 kDa (alpha chain) in wild-type MCF7 cell lysates with no signal observed at this size in IGF1R knockout cell line. To generate this image, wild-type and IGF1R knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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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![Western blot - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)](https://www.abcam.com/ps/products/263/ab263907/Images/ab263907-450233-anti-igf1-receptor-antibody-epr23027-80-western-blot-hela-lysates.jpg)
Western blot - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)All lanes : Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : IGF1R knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 154 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab263907 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab263907 was shown to react with IGF1 Receptor in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264801 (knockout cell lysate ab256951) was used. Wild-type HeLa and IGF1R knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab263907 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)](https://www.abcam.com/ps/products/263/ab263907/Images/ab263907-1-anti-igf1-receptor-antibody-epr23027-80-immunocytochemistry-mcf7-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)Immunofluorescent analysis of 100% Methanol-fixed MCF7 (human breast adenocarcinoma epithelial cell) cells labelling IGF1 Receptor with ab263907 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in MCF7 cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
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![Flow Cytometry - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)](https://www.abcam.com/ps/products/263/ab263907/Images/ab263907-2-anti-igf1-receptor-antibody-epr23027-80-flowcytometry-mcf7-human.jpg)
Flow Cytometry - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)Flow cytometric analysis MCF7 (human breast adenocarcinoma epithelial cell) cells labelling IGF1 Receptor with ab263907 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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![Western blot - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)](https://www.abcam.com/ps/products/263/ab263907/Images/ab263907-4-anti-igf1-receptor-antibody-epr23027-80-western-blot-human.jpg)
Western blot - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)All lanes : Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907) at 1/1000 dilution
Lane 1 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 154 kDa
Observed band size: 130,200 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 10 seconds.
The expression profile & molecular weight observed is consistent with what has been described in the literature (PMID: 21807868, 28591735).
Note: the bands larger than 200kDa are Pro-IGF1R.
Low expression cell line: MDA-MB-231 (PMID: 28591735).
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![Immunoprecipitation - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)](https://www.abcam.com/ps/products/263/ab263907/Images/ab263907-3-anti-igf1-receptor-antibody-epr23027-80-immunoprecipitation-mcf7-human.jpg)
Immunoprecipitation - Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)IGF1 Receptor was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab263907 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263907 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2: ab263907 IP in MCF7 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263907 in MCF7 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 min.
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![Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)](https://www.abcam.com/ps/products/263/ab263907/Images/ab263907-5-benefits-of-recombinant-antibodies.png)
Anti-IGF1 Receptor antibody [EPR23027-80] (ab263907)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab263907 has been referenced in 2 publications.
- Ding Y et al. Circular RNA midline-1 (circMID1) promotes proliferation, migration, invasion and glycolysis in prostate cancer. Bioengineered 13:6293-6308 (2022). WB ; Human . PubMed: 35212614
- Zhang H et al. miR-520b Inhibits IGF-1R to Increase Doxorubicin Sensitivity and Promote Cell Apoptosis in Breast Cancer. Yakugaku Zasshi 141:415-426 (2021). PubMed: 33116033