EGFR Antibody - #AF6043

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製品: EGFR Antibody
カタログ: AF6043
タンパク質の説明: Rabbit polyclonal antibody to EGFR
アプリケーション: WB IHC IF/ICC
Cited expt.: WB, IHC, IF/ICC
反応性: Human, Mouse, Rat
予測: Pig, Bovine, Sheep, Rabbit
分子量: 134kDa; 134kD(Calculated).
ユニプロット: P00533
RRID: AB_2834971

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製品説明

ソース:
Rabbit
アプリケーション:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(92%), Bovine(92%), Sheep(92%), Rabbit(92%)
クローナリティ:
Polyclonal
特異性:
EGFR Antibody detects endogenous levels of total EGFR.
RRID:
AB_2834971
引用形式: Affinity Biosciences Cat# AF6043, RRID:AB_2834971.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

Avian erythroblastic leukemia viral (v erb b) oncogene homolog; Cell growth inhibiting protein 40; Cell proliferation inducing protein 61; EGF R; EGFR; EGFR_HUMAN; Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog); Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian); Epidermal growth factor receptor; erb-b2 receptor tyrosine kinase 1; ERBB; ERBB1; Errp; HER1; mENA; NISBD2; Oncogen ERBB; PIG61; Proto-oncogene c-ErbB-1; Receptor tyrosine protein kinase ErbB 1; Receptor tyrosine-protein kinase ErbB-1; SA7; Species antigen 7; Urogastrone; v-erb-b Avian erythroblastic leukemia viral oncogen homolog; wa2; Wa5;

免疫原

免疫原:

A synthesized peptide derived from human EGFR, corresponding to a region within C-terminal amino acids.

Uniprot:

P00533(EGFR_HUMAN) >>Visit HPA database.

遺伝子(ID):
EGFR(1956)
発現特異性:
P00533 EGFR_HUMAN:

Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.

タンパク質の説明:
EGFR is a receptor tyrosine kinase. Receptor for epidermal growth factor (EGF) and related growth factors including TGF-alpha, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30 and vaccinia virus growth factor. Is involved in the control of cell growth and differentiation.
タンパク質配列:
MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFEDHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENLQIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQWRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGRCRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNPEGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGIGIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTPSGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSPKANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQYLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHAEGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKGERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHLPSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVACIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQNPVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQISLDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
92
Bovine
92
Sheep
92
Rabbit
92
Xenopus
69
Horse
0
Dog
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

研究背景

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, AREG, 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. Positively regulates cell migration via interaction with CCDC88A/GIV which retains EGFR at the cell membrane following ligand stimulation, promoting EGFR signaling which triggers cell migration. Plays a role in enhancing learning and memory performance (By similarity).

Isoform 2 may act as an antagonist of EGF action.

(Microbial infection) Acts as a receptor for hepatitis C virus (HCV) in hepatocytes and facilitates its cell entry. Mediates HCV entry by promoting the formation of the CD81-CLDN1 receptor complexes that are essential for HCV entry and by enhancing membrane fusion of cells expressing HCV envelope glycoproteins.

PTMs:

Phosphorylated on Tyr residues in response to EGF. 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 (By similarity).

Palmitoylated on Cys residues by ZDHHC20. Palmitoylation inhibits internalization after ligand binding, and increases the persistence of tyrosine-phosphorylated EGFR at the cell membrane. Palmitoylation increases the amplitude and duration of EGFR signaling.

Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197.

細胞の位置付け:

Cell membrane>Single-pass type I membrane protein. Endoplasmic reticulum membrane>Single-pass type I membrane protein. Golgi apparatus membrane>Single-pass type I membrane protein. Nucleus membrane>Single-pass type I membrane protein. Endosome. Endosome membrane. Nucleus.
Note: In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER (PubMed:20674546). Endocytosed upon activation by ligand (PubMed:2790960, PubMed:17182860, PubMed:27153536). Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF) (PubMed:20551055).

Secreted.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.

タンパク質ファミリー:

Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.

研究領域

· Cellular Processes > Transport and catabolism > Endocytosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Adherens junction.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Gap junction.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (View pathway)

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Calcium signaling pathway.   (View pathway)

· Environmental Information Processing > Signaling molecules and interaction > Cytokine-cytokine receptor interaction.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Infectious diseases: Bacterial > Epithelial cell signaling in Helicobacter pylori infection.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Endometrial cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Glioma.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Bladder cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Choline metabolism in cancer.   (View pathway)

· Organismal Systems > Endocrine system > Estrogen signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

参考文献

1). Enhancement of CAR-T cell activity against cholangiocarcinoma by simultaneous knockdown of six inhibitory membrane proteins. Cancer Communications, 2023 (PubMed: 37282786) [IF=16.2]

Application: IHC    Species: Human    Sample:

FIGURE 1 EGFR‐CAR T cells and B7H3‐CAR T cells exhibited general antitumor activity in cholangiocarcinoma cell lines. (A‐B) Detection of tumor tissue antigens collected from patients with cholangiocarcinoma by IHC staining. The left and right panels of A displayed representative EGFR and B7H3 expression in normal, para‐carcinoma area, and tumor zone with enlarged view (5x, scale bar = 200 μm; 20x, scale bar = 50 μm). Statistical analysis of EGFR and B7H3 antigens scores in normal, para‐carcinoma and carcinoma tissues (B, n = 20). (C) Schematic representation of the lentiviral vectors carrying an EGFR‐specific 3rd generation CAR moiety (EGFR‐CAR) and a B7H3‐specific 3rd generation CAR moiety (B7H3‐CAR). (D) Immunoblotting analysis of EGFR‐CAR and B7H3‐CAR expression were performed with anti‐flag tag. The positive control was GP120‐CAR. (E‐F) Detection cytotoxic activities of the EGFR‐CAR‐T cells and the B7H3‐CAR‐T cells with TFK‐1 cells and HuCCT1 cells by LDH assay. The negative control was Jurkat cells without EGFR and B7H3 expression (n = 3, healthy donors). Experiments were performed independently at least 3 times. One‐way ANOVA was used in Tukey's multiple comparison test. Experimental data are expressed as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Abbreviations: CAR, chimeric antigen receptor T; EGFR, epidermal growth factor receptor; B7H3, B7 homolog 3; GP120, gp120 antigen; IHC, immunohistochemistry; LDH, lactate dehydrogenase; SEM, standard error of the mean.
2). Foxq1 promotes metastasis of nasopharyngeal carcinoma by inducing vasculogenic mimicry via the EGFR signaling pathway. Cell Death & Disease, 2021 (PubMed: 33875643) [IF=8.1]

Application: WB    Species: mouse    Sample: 5–8F cells and CNE1

Fig. 3| Foxq1 increases the expression of EGFR, while Erlotinib inhibits Foxq1-induced VM formation. A, B qRT-PCR was used to assess the expression of predicted genes in 5–8F cells that overexpressed miR-124 or had a Foxq1 knockdown. C–F qRT-PCR (left) and western blot (right) of EGFR expression in Foxq1 dysregulated 5–8F, and CNE1.

Application: IHC    Species: mouse    Sample: NPC tissues

Fig. 3| Foxq1 increases the expression of EGFR, while Erlotinib inhibits Foxq1-induced VM formation. A, B qRT-PCR was used to assess the expression of predicted genes in 5–8F cells that overexpressed miR-124 or had a Foxq1 knockdown. C–F qRT-PCR (left) and western blot (right) of EGFR expression in Foxq1 dysregulated 5–8F, and CNE1.I Region showing typical expression of Foxq1 and EGFR in NPC tissues; Scale bars represent 50 μm.

Application: IF/ICC    Species: mouse    Sample: 5–8F cells

Fig. 7| MiR-124 inhibits the EGFR signaling pathway and VM formation, that could be rescued by Foxq1 expression. E Immunofluorescence staining of Foxq1, EGFR, VE-cadherin, MMP2 and MMP9 in 5–8F cells that overexpressed miR-124, control or simultaneously Foxq1 and miR-124, respectively; scale bars represent 50μm. qRT-PCR (F) and western blot (G) were used to monitor the expression of EGFR signaling pathway and VM-related genes in 5–8F cells that overexpressed miR-124, control or simultaneously Foxq1 and miR-124, respectively.Data are presented as mean ± SD of three independent experiments.
3). Higenamine alleviates allergic rhinitis by activating AKT1 and suppressing the EGFR/JAK2/c-JUN signaling. PHYTOMEDICINE, 2021 (PubMed: 33945919) [IF=6.7]

Application: WB    Species: Human    Sample: HNEpCs

Fig. 8. The effects of higenamine (HG) on potential targets in histamine-induced HNEpCs. (A-J) Levels of protein expression of AKT1, p-AKT1, EGFR, p-EGFR, c-Jun, p-c-Jun, iNOS, JAK2, and p-JAK2 were determined by Western blotting, β-actin was used as an internal control. Data are expressed as means ± SD of three experiments. ## p < 0.01, and # p < 0.05 compared with the control group. ** p < 0.01, and * p < 0.05 compared with the histamine group.
4). Resistance to anti-HER2 therapy associated with the TSC2 nonsynonymous variant c.4349 C > G (p.Pro1450Arg) is reversed by CDK4/6 inhibitor in HER2-positive breast cancer. npj Breast Cancer, 2023 (PubMed: 37160904) [IF=5.9]

Application: WB    Species: Human    Sample: BT474 cell

Fig. 3 Expression of proteins involved in the HER2/AKT/mTOR and cyclinD1/CDK4 signaling pathways in BT474 cell lines treated with lapatinib and/or palbociclib. a Western blot analysis of proteins involved in the HER2/AKT/mTOR signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. b Western blot analysis of proteins involved in the cyclinD1/CDK4 signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. c p-HER2 expression levels under treatment with different drugs in the same cell. d p-HER2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. e p-TSC2 expression levels under treatment with different drugs in the same cell. f p-TSC2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. g p-mTOR expression levels under treatment with different drugs in the same cell. h p-mTOR expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. i p-P70S6K expression levels under treatment with different drugs in the same cell. j p-P70S6K expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. Data are shown as mean ± SD; One-way ANOVA was used to analyze the data in (d, f, h, j). Error bars are SEM.
5). Ginsenoside-Rh2 Promotes Functional Recovery after Spinal Cord Injury by Enhancing TFEB-Mediated Autophagy. Journal of agricultural and food chemistry, 2024 (PubMed: 38907713) [IF=5.7]
6). TIAM2 Contributes to Osimertinib Resistance, Cell Motility, and Tumor-Associated Macrophage M2-like Polarization in Lung Adenocarcinoma. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2022 (PubMed: 36142328) [IF=5.6]
7). Increasing the Hindgut Carbohydrate/Protein Ratio by Cecal Infusion of Corn Starch or Casein Hydrolysate Drives Gut Microbiota-Related Bile Acid Metabolism To Stimulate Colonic Barrier Function. mSystems, 2020 (PubMed: 32487741) [IF=5.0]

Application: WB    Species: Human    Sample: Colon

FIG 5 (A to C) Effects of selected bile acid on barrier function (A)-, bile acid receptor (B)-, and epithelial barrier function regulation (C)-related gene expression of Caco-2 cells (n  3). (D) Effects of selected bile acid on the relative protein levels of ZO-1, Src, and EGFR (n  3). GAPDH was used as an internal control. (E) Effects of selected bile acid and EGFR-specific inhibitor (AG-1478) on the relative protein levels of ZO-1, Src, and EGFR (n  3). GAPDH was used as an internal control. (F) Effects of selected bile acid and Src family kinase inhibitor (PP-2) on the relative protein levels of ZO-1 and Src (n  3). GAPDH was used as an internal control. Data are presented as mean  SEM. *, compared with control, P  0.05; **, compared with control, P  0.01; ***, compared with control, P  0.001; , compared with DCA group, P  0.05; , compared with LCA group, P  0.05. CA, cholic acid; DCA, deoxycholic acid; LCA, lithocholic acid; OCLD, occludin; ZO-1, zonula occludens 1; FXR, farnesoid X receptor; TGR5, G-protein-coupled receptor; EGFR, epidermal growth factor receptor; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PI3K, phosphatidylinositol 3-kinase; RHOA, ras homolog gene family, member A; MLCK, myosin light chain kinase.
8). Processed product (Pinelliae Rhizoma Praeparatum) of Pinellia ternata (Thunb.) Breit. Alleviates the allergic airway inflammation of cold phlegm via regulation of PKC/EGFR/MAPK/PI3K-AKT signaling pathway. JOURNAL OF ETHNOPHARMACOLOGY, 2022 (PubMed: 35688257) [IF=4.8]
9). Jiawei Yanghe Decoction suppresses breast cancer by regulating immune responses via JAK2/STAT3 signaling pathway. JOURNAL OF ETHNOPHARMACOLOGY, 2023 (PubMed: 36933872) [IF=4.8]
10). Systematic analysis of the mechanism of hydroxysafflor yellow A for treating ischemic stroke based on network pharmacology technology. European Journal of Pharmacology, 2021 (PubMed: 34302817) [IF=4.2]

Application: IHC    Species: Rat    Sample: hippocampal

Fig. 12. Immunohistochemical staining and statistical results. The expression levels of positive cells in the hippocampal CA1 area of rats were observed under an optical microscope at a magnification of × 200. Sham versus I/R, #P < 0.05, ##P < 0.01; HSYA versus I/R, **P < 0.01.

Application: WB    Species: Rat    Sample: hippocampal

Fig. 13. Western blot results for the detection of EGF receptor, HIF1A and NOS3. Sham versus I/R, #P < 0.05; HSYA versus I/R, **P < 0.01; a–b EGF receptor; c–d HIF1A; e–f NOS3.
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