Anti-EGFR (phospho Y1068) 抗体 (ab5644)
Key features and details
- Rabbit polyclonal to EGFR (phospho Y1068)
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-EGFR (phospho Y1068) antibody
EGFR 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to EGFR (phospho Y1068) -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, WBmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide corresponding to Human EGFR (phospho W1068).
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ポジティブ･コントロール
- ウェスタン・ブロット用ポジティブコントロールRecombinant human EGFR protein
- WB: A-431 and A549 whole cell lysate treated with EGF (200 ng/mL for 10 minutes). ICC: A-431 cells treated with 200 ng/mL of EGF for 10 minutes.
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特記事項
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
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
バッファー
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA -
Concentration information loading... -
精製度
Immunogen affinity purified -
特記事項（精製）
The antibody has been negatively preadsorbed using (i) a non phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated epidermal growth factor receptor (EGFR), and (ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphotyrosine, irrespective of the sequence. The final product is generated by affinity chromatography using an EGFR-derived peptide that is phosphorylated at tyrosine 1068. -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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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ab5644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 |
|---|---|---|
| ICC/IF |
1/100.
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| WB | (5) |
1/1000. Detects a band of approximately 185 kDa.
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| 特記事項 |
|---|
|
ICC/IF
1/100. |
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WB
1/1000. Detects a band of approximately 185 kDa. |
ターゲット情報
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機能
Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules. May also activate the NF-kappa-B signaling cascade. Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin.
Isoform 2 may act as an antagonist of EGF action. -
組織特異性
Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers. -
関連疾患
Lung cancer
Inflammatory skin and bowel disease, neonatal, 2 -
配列類似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
翻訳後修飾
Phosphorylation at Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Phosphorylation at Thr-678 and Thr-693 by PRKD1 inhibits EGF-induced MAPK8/JNK1 activation. Dephosphorylation by PTPRJ prevents endocytosis and stabilizes the receptor at the plasma membrane. Autophosphorylation at Tyr-1197 is stimulated by methylation at Arg-1199 and enhances interaction with PTPN6. Autophosphorylation at Tyr-1092 and/or Tyr-1110 recruits STAT3. Dephosphorylated by PTPN1 and PTPN2.
Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197. -
細胞内局在
Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF). - Information by UniProt
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参照データベース
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
- Unigene: 605083 Human
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別名
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
- Cell growth inhibiting protein 40 antibody
- Cell proliferation inducing protein 61 antibody
see all
画像
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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody (ab5644)
Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR with ab5644 at 1/100 dilution, followed by Goat anti-rabbit IgG (H+L) Superclonal, Alexa Fluor® 488 conjugate was used as the secondary antibody at 1/2000 dilution (Panel a). Nuclei (Panel b) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c) was stained with Rhodamine Phalloidin. Panel (d) represents the merged image showing membrane localization. Panel (e) represents cells treated with antagonist, Afatinib (1µM for 6hrs) followed by EGF (200 ng/ml for 10 minutes), showing no Phospho-EGFR staining. Panel (f) shows untreated cells with no signal. Panel (g) represents control cells with no primary antibody to assess background.
The cells (in 70% confluent log phase treated with 200ng/ml of EGF for 10 minutes) were fixed with 4% paraformaldehyde for 10 minutes; permeabilized with 0.1% Triton X-100 for 10 minutes and blocked with 1% BSA for hour at room temperature. The images were captured at 60X magnification.
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Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)All lanes : Anti-EGFR (phospho Y1068) antibody (ab5644) at 1/1000 dilution
Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate with skimmed milk
Lane 2 : A431 whole cell lysate treated with EGF (200 ng/mL for 10 minutes) with skimmed milk
Lane 3 : A431 whole cell lysate treated with Gefitinib followed by EGF (1uM for 16 hours, 200 ng/mL for 10 minutes) with skimmed milk
Lane 4 : A431 whole cell lysate treated with Afatinib followed by EGF (0.5 uM for 6 hours, 200 ng/mL for 10 minutes) with skimmed milk
Lane 5 : A549 (Human lung carcinoma cell line) whole cell lysate with skimmed milk
Lane 6 : A549 whole cell lysate treated with EGF (200 ng/mL for 10 minutes) with skimmed milk
Lane 7 : A549 whole cell lysate treated with Afatinib followed by EGF (0.5 uM for 6 hours, 200 ng/mL for 10 minutes) with skimmed milk
Lysates/proteins at 30 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat anti-Rabbit IgG Superclonal Secondary Antibody, HRP at 1/4000 dilutionWestern blot analysis using ab5644 shows increased expression of proteins phosphorylated at the tyrosine residues in A-431 and A549 cell lines upon EGF treatment and pre-treatment with EGFR-antagonists, Gefitinib and Afatinib. This results in inhibition of Phospho-EGFR in A-431 and A549 cell lines
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Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)This image is courtesy of an Abreview submitted by Mr Samir NuseibehAll lanes : Anti-EGFR (phospho Y1068) antibody (ab5644) at 1/200 dilution
Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate - not treated
Lane 2 : A431 whole cell lysate - 100ng/ml EGF for 1 minute
Lane 3 : A431 whole cell lysate - 100ng/ml EGF for 2.5 minutes
Lane 4 : A431 whole cell lysate - 100ng/ml EGF for 5 minutes
Lane 5 : A431 whole cell lysate - 100ng/ml EGF for 10 minutes
Lane 6 : A431 whole cell lysate - 100ng/ml EGF for 20 minutes
Lane 7 : A431 whole cell lysate - 100ng/ml EGF for 40 minutes
Secondary
All lanes : HRP conjugated Goat anti-rabbit antibody
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 2 minutes
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (43)
ab5644 は 43 報の論文で使用されています。
- Aldonza MBD et al. Chemotherapy confers a conserved secondary tolerance to EGFR inhibition via AXL-mediated signaling bypass. Sci Rep 11:8016 (2021). PubMed: 33850249
- Mizuta H et al. Gilteritinib overcomes lorlatinib resistance in ALK-rearranged cancer. Nat Commun 12:1261 (2021). PubMed: 33627640
- Fernandes Neto JM et al. Multiple low dose therapy as an effective strategy to treat EGFR inhibitor-resistant NSCLC tumours. Nat Commun 11:3157 (2020). PubMed: 32572029
- Sun C et al. FGL1 regulates acquired resistance to Gefitinib by inhibiting apoptosis in non-small cell lung cancer. Respir Res 21:210 (2020). PubMed: 32778129
- Ma ZJ et al. LncZEB1-AS1 regulates hepatocellular carcinoma bone metastasis via regulation of the miR-302b-EGFR-PI3K-AKT axis. J Cancer 11:5118-5128 (2020). PubMed: 32742459