Anti-IGF1 Receptor 抗体 [EPR23027-204] (ab263903)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23027-204] to IGF1 Receptor
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-IGF1 Receptor antibody [EPR23027-204]
IGF1 Receptor 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR23027-204] to IGF1 Receptor -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-Pmore details
適用なし: Flow Cyt,ICC/IF or IP -
種交差性
交差種: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: A431, HepG2, HeLa and MDA-MB-231 lysates. IHC-P: Human breast carcinoma, Human prostatic hyperplasia and Human bladder carcinoma tissues.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR23027-204 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab263903の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
1/1000. Detects a band of approximately 130, 200 kDa (predicted molecular weight: 154 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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特記事項 |
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WB
1/1000. Detects a band of approximately 130, 200 kDa (predicted molecular weight: 154 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ターゲット情報
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機能
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. -
組織特異性
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. -
関連疾患
Insulin-like growth factor 1 resistance -
配列類似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. Insulin receptor subfamily.
Contains 4 fibronectin type-III domains.
Contains 1 protein kinase domain. -
翻訳後修飾
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. -
細胞内局在
Cell membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 3480 Human
- Omim: 147370 Human
- SwissProt: P08069 Human
- Unigene: 643120 Human
- Unigene: 714012 Human
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別名
- CD221 antibody
- CD221 antigen antibody
- IGF 1 receptor antibody
see all
画像
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All lanes : Anti-IGF1 Receptor antibody [EPR23027-204] (ab263903) at 1/1000 dilution
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : IGF1R knockout HCT 116 cell lysate
Lane 3 : Wild-type MCF7 ab290784 cell lysate
Lane 4 : IGF1R knockout MCF7 ab287507 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 154 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?Anti-IGF1R antibody [EPR23027-204] (ab263903) 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, ab263903 was shown to bind specifically to IGF1R. A band was observed at 105 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in IGF1R knockout cell line. To generate this image, wild-type and IGF1R knockout HCT 116 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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All lanes : Anti-IGF1 Receptor antibody [EPR23027-204] (ab263903) 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 [EPR23027-204] 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, ab263903 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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Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling IGF1 Receptor with ab263903 at 1/500 dilution (1.24 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Membranous and cytoplasmic staining on human prostatic hyperplasia (PMID: 20710042). The section was incubated with ab263903 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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All lanes : Anti-IGF1 Receptor antibody [EPR23027-204] (ab263903) 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 : Rabbit polyclonal to GNAT2 (ab97501) at 1/100000 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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Immunohistochemical analysis of paraffin-embedded Human bladder carcinoma tissue labeling IGF1 Receptor with ab263903 at 1/500 dilution (1.24 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Membranous and cytoplasmic staining on human bladder carcinoma (PMID: 20710042). The section was incubated with ab263903 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling IGF1 Receptor with ab263903 at 1/500 dilution (1.24 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Membranous and cytoplasmic staining on human breast carcinoma (PMID: 21057462). The section was incubated with ab263903 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (3)
ab263903 は 3 報の論文で使用されています。
- Xu W et al. Immunogenomic Characteristics of Cell-Death-Associated Genes with Prognostic Implications in Bladder Cancer. Front Immunol 13:909324 (2022). PubMed: 35898507
- Ortega MA et al. Abnormal proinflammatory and stressor environmental with increased the regulatory cellular IGF-1/PAPP-A/STC and Wnt-1/β-Catenin canonical pathway in placenta of women with Chronic venous Disease during Pregnancy. Int J Med Sci 18:2814-2827 (2021). PubMed: 34220309
- Ortega MA et al. Chronic Venous Disease Patients Showed Altered Expression of IGF-1/PAPP-A/STC-2 Axis in the Vein Wall. Biomed Res Int 2020:6782659 (2020). PubMed: 33381575