Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

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Overview

  • Product name

    Anti-IGF1 Receptor antibody [EPR19322]
    See all IGF1 Receptor primary antibodies

  • Description

    Rabbit monoclonal [EPR19322] to IGF1 Receptor

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Mouse IGF1 Receptor aa 700-950. The exact sequence is proprietary.
    Database link: Q60751

  • Positive control

    • WB: C2C12, HeLa, 293, MCF7, C6, and NIH/3T3 whole cell lysates; Mouse brain and spleen lysates; Rat brain lysate. ICC/IF: C2C12 and C6 cells. Flow Cyt: C2C12 cells. IP: C2C12 whole cell lysate.

  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab182408 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 100,200 kDa (predicted molecular weight: 156 kDa).
Flow Cyt 1/80.
ICC/IF 1/1000.
IP 1/50.

Target

  • 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
    modifications

    Autophosphorylated 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

  • Database links

    see all

  • Alternative names

    • CD221 antibody
    • CD221 antigen antibody
    • IGF 1 receptor antibody
    • IGF 1R antibody
    • IGF I receptor antibody
    • IGF-I receptor antibody
    • Igf1r antibody
    • IGF1R_HUMAN antibody
    • IGFIR antibody
    • IGFIRC antibody
    • IGFR antibody
    • Insulin like growth factor 1 receptor antibody
    • Insulin like growth factor 1 receptor precursor antibody
    • Insulin-like growth factor 1 receptor beta chain antibody
    • Insulin-like growth factor I receptor antibody
    • JTK13 antibody
    • MGC142170 antibody
    • MGC142172 antibody
    • MGC18216 antibody
    • Soluble IGF1R variant 1 antibody
    • Soluble IGF1R variant 2 antibody
    see all

Images

  • Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

    Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
    Lane 2: IGF1R knockout  HAP1 whole cell lysate (40 µg)
    Lane 3: HeLa whole cell lysate (40 µg)

    Lanes 1 - 3 Merged signal (red and green). Green - ab182408 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab182408 was shown to specifically recognize IGF1R in wild-type HAP1 cells along with additional cross-reactve bands. No band was observed when IGF1R knockout samples were examined. Wild-type and IGF1R knockout samples were subjected to SDS-PAGE.  Ab182408 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.         

  • Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cell line) cells labeling IGF1 Receptor with ab182408 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on C6 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:

    -ve control 1: ab182408 at 1/1000 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Lanes 1-4 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/2000 dilution
    Lanes 5-6 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/1000 dilution

    Lane 1 : C2C12 (Mouse myoblast cell line) whole cell lysate at 20 µg
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
    Lane 3 : 293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
    Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate at 10 µg
    Lane 6 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 156 kDa
    Observed band size: 100,200 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 5 seconds; Lane 2-4: 3 minutes; Lane 5/6: 2 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 11402025).

  • Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (Mouse myoblast cell line) cells labeling IGF1 Receptor with ab182408 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on C2C12 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:

    -ve control 1: ab182408 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Lane 1 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/1000 dilution
    Lanes 2-3 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse spleen lysate
    Lane 3 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 156 kDa
    Observed band size: 100,200 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 10 seconds; Lane 2: 3 minutes; Lane 3: 1 minute.

    The expression profile observed is consistent with what has been described in the literature (PMID: 11402025).

  • Flow Cytometry - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Flow Cytometry - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

    Flow cytometric analysis of 4% paraformaldehyde-fixed C2C12 (Mouse myoblast cell line) cells labeling IGF1 Receptor with ab182408 at 1/80 dilution (red) compared with a Rabbit IgG,monoclonal[EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

     

     

  • Immunoprecipitation - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)
    Immunoprecipitation - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

    IGF1 Receptor was immunoprecipitated from 1mg of C2C12 (Mouse myoblast cell line) whole cell lysate with ab182408 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab182408 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: C2C12 whole cell lysate, 10µg (Input).

    Lane 2: ab182408 IP in C2C12 whole cell lysate.

    Lane 3: Rabbit  IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab182408 in C2C12 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

References

This product has been referenced in:

  • Chen L  et al. microRNA-628 inhibits the proliferation of acute myeloid leukemia cells by directly targeting IGF-1R. Onco Targets Ther 12:907-919 (2019). Read more (PubMed: 30774377) »
  • Jiang W  et al. Wentilactone A induces cell apoptosis by targeting AKR1C1 gene via the IGF-1R/IRS1/PI3K/AKT/Nrf2/FLIP/Caspase-3 signaling pathway in small cell lung cancer. Oncol Lett 16:6445-6457 (2018). Read more (PubMed: 30405782) »
See all 8 Publications for this product

Publishing research using ab182408? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Western blot abreview for Anti-IGF1 Receptor antibody [EPR19322]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Sheep Tissue lysate - whole (fetal skeletal muscle)
Gel Running Conditions
Non-reduced Denaturing (10)
Loading amount
50 µg
Specification
fetal skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Camila Sandoval Torres

Verified customer

Submitted Dec 17 2018

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