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P egfr

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P-EGFR is an antibody that detects the phosphorylated form of the epidermal growth factor receptor (EGFR). It is used in research applications to study the activation and signaling of the EGFR pathway.

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Lab products found in correlation

β actin, by Santa Cruz Biotechnology (7 mentions) Gapdh, by Santa Cruz Biotechnology (7 mentions) P erk, by Santa Cruz Biotechnology (6 mentions) P akt, by Santa Cruz Biotechnology (6 mentions) P akt, by Cell Signaling Technology (4 mentions) β actin, by Merck Group (3 mentions) β catenin, by Cell Signaling Technology (2 mentions) Vascular endothelial growth factor (vegf), by Abcam (2 mentions) Erk1 2, by Cell Signaling Technology (2 mentions) P erk1 2, by Cell Signaling Technology (2 mentions) E cadherin, by Cell Signaling Technology (2 mentions) P erbb2, by Santa Cruz Biotechnology (2 mentions) P s6k, by Cell Signaling Technology (2 mentions) β actin, by Cell Signaling Technology (2 mentions) P fak, by Abcam (2 mentions) Stat3, by Santa Cruz Biotechnology (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Cyclin d1, by Santa Cruz Biotechnology (2 mentions) P erk1 2, by Santa Cruz Biotechnology (2 mentions) Imagequant las 500 analyzer, by GE Healthcare (2 mentions)

Spelling variants of this product

P-EGFR (Y1068) (3 citations) P-EGFR (Tyr 1173) (3 citations) P-EGFR (Tyr845) (2 citations)

29 protocols using p egfr

1

Comprehensive Protein Expression Analysis

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Whole-cell lysates were prepared as previously described55 (link). Antibodies specific for p53 (1:2000, sc-126), p-EGFR (1:1000, Tyr1173, sc-101668), EGFR (1:2000, sc-373749), p-Erk (1:1000, Thr202/Tyr204, sc-16982), Erk (1:3000, sc-94), Akt (1:3000, sc-5298), AXL (1:1000, sc-1096), E-cadherin (1:1000, sc-71008), EpCAM (1:1000, sc-71059), desmoplakin (1:1000, sc-390975), cytokeratin-8/18 (1:1000, sc-70939) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); those for p-Akt (1:1000, Ser473, #4060), β-catenin (1:1000, #8480), p-AXL (1:1000, #4060), and vimentin (1:1000, #5741) were obtained from Cell Signaling Technology (Beverly, MA, USA). To assess the level of p-AXL, lysates were immunoprecipitated with an anti-AXL antibody and immunoblotted with an anti-phosphotyrosine (p-Tyr, 1:1000, sc-7020, Santa Cruz) antibody. The immunoblotting is representative of three independent experiments.
Jung S., Kim D.H., Choi Y.J., Kim S.Y., Park H., Lee H., Choi C.M., Sung Y.H., Lee J.C, & Rho J.K. (2021). Contribution of p53 in sensitivity to EGFR tyrosine kinase inhibitors in non-small cell lung cancer. Scientific Reports, 11, 19667.
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2

Myxoid Liposarcoma EGFR Activation

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Myxoid liposarcoma cells were stimulated with macrophage-conditioned medium (M-CM: RAW-CM and U937-CM) or HB-EGF 20 ng ml−1 for 5 min. To inhibit the stimulation, various concentrations of gefitinib and anti-HB-EGF antibody were added for 2 h before the stimulation. After stimulation, the cells were lysed using CelLytic (Sigma-Aldrich) with a protease and phosphatase inhibitor cocktail (Complete Mini, PhosSTOP; Roche Diagnostics, Mannheim, Germany). Western blot analysis was performed as described previously (Fujiwara-Okada et al, 2013 (link); Iida et al, 2013 (link)) with the following primary antibodies: pEGFR (1 : 500), EGFR (1 : 1000), and β-actin (1 : 2000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).
Nabeshima A., Matsumoto Y., Fukushi J., Iura K., Matsunobu T., Endo M., Fujiwara T., Iida K., Fujiwara Y., Hatano M., Yokoyama N., Fukushima S., Oda Y, & Iwamoto Y. (2015). Tumour-associated macrophages correlate with poor prognosis in myxoid liposarcoma and promote cell motility and invasion via the HB-EGF-EGFR-PI3K/Akt pathways. British Journal of Cancer, 112(3), 547-555.
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3

Multimarker Immunohistochemistry Analysis

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Tissues were fixed in 4% paraformaldehyde, sent for sectioning, and assessed with an IHC kit (ZSGB-BIO, Beijing, China). Slides were incubated at 4°C overnight (16−20 h) with antibodies reactive with Ki67 (#ab16667; Abcam), F4/80 (#ab100790; Abcam), CD163 (#sc58965; Santa Cruz), VEGF (#ab32152; Abcam), VEGFR (#ab32152; Abcam), CD34 (#ab81289; Abcam), EGFR (#sc373746; Santa Cruz), and p-EGFR (#sc-377547; Santa Cruz). Diaminobenzidine (K176810E; ZSGB-BIO, China) was used for target protein staining and H&E for nuclear staining. Images were photographed at 400× with a microscope.
He S., Tian S., He X., Le X., Ning Y., Chen J., Chen H., Mu J., Xu K., Xiang Q., Wu Y., Chen J, & Xiang T. (2021). Multiple targeted self-emulsifying compound RGO reveals obvious anti-tumor potential in hepatocellular carcinoma. Molecular Therapy Oncolytics, 22, 604-616.
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4

MTT Assay and Signaling Pathway Analysis

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3-(4,5-Dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI) and other chemicals were purchased from Sigma (St. Louis, Missouri, USA). The specific anti-SHP1 short interfering RNA (siRNA) and nonspecific control siRNA sequences were purchased from GenePharma (Shanghai, China). Commercial antibodies to the following antigens were as follows: SHP-1, EGFR, p-GSK3β (Ser9), Cyclin D1, and c-Myc (Epitomics, Burlingame, USA); p-EGFR, β-actin, GAPDH, histone H3, and EGFR (Santa Cruz Biotechnology, California, USA); h-Ras, p-Erk1/2, Erk1/2, β-catenin, Snail, E-cadherin, and N-cadherin (Cell Signaling Technology, Massachusetts, USA); k-Ras, GSK3β (Proteintech Group, Chicago, USA); and SHP-1 (Abcam, Cambridge, UK).
Geng Q., Xian R., Yu Y., Chen F, & Li R. (2021). SHP-1 acts as a tumor suppressor by interacting with EGFR and predicts the prognosis of human breast cancer. Cancer Biology & Medicine, 19(4), 468-485.
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5

Protein Expression Analysis via Western Blotting

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Protein extraction and Western blotting were performed as previously described (24 (link)). Primary antibodies against IL17RD, FLAG, pAKT, pERK, pERBB2 EGFR, pEGFR, and beta actin were used with a dilution of 1:500 for IL17RD (R and D systems), 1:1000 for FLAG (Clontech, Mountain View, CA),1:500 pAKT (Cell Signaling Technology, Danvers, MA), 1:500 for EGFR (Cell Signaling Technology), 1:100 for pEGFR (Santa Cruz Biotechnology, Dallas, TX), 1:2500 for pERK (Santa Cruz Biotechnology), 1:200 for pERBB2 (Santa Cruz Biotechnology) and 1:5000 for beta actin (Sigma Aldrich, St. Louis, MO). Mean expression between treatment group was compared using a Student’s t-test.
Pekow J., Meckel K., Dougherty U., Huang Y., Chen X., Almoghrabi A., Mustafi R., Ayaloglu-Butun F., Deng Z., Haider H.I., Hart J., Rubin D.T., Kwon J.H, & Bissonnette M. (2017). miR-193a-3p is a key tumor suppressor in ulcerative colitis-associated colon cancer and promotes carcinogenesis through up-regulation of IL17RD. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(17), 5281-5291.
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6

Comprehensive Protein Expression Analysis

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Antibodies against the following proteins were used: PLD2 (sc-515744), BAX (sc-7480), XIAP (sc-55550), Bim (sc-374358), VEGF (sc-7269), EGFR (sc-373746), p-EGFR (sc-81488), AKT (sc-81434), p-AKT (sc-377556), PKCζ (sc-17781), pPKC (sc-12894R), PKCδ (sc-8402), p-PKCδ (sc-377560) and α-tubulin (sc-8035, Santa Cruz Biotechnology, Dallas, TX, USA), acetyl-Histone 4 (06-866, EMD Millipore, Burlington, MA, USA), ERK (#9102), p-ERK (#9101S), JNK (#9252), p-JNK (#4668), IκBα (#4814), p-IκBα (#2859), Src (#2109), p-Src (#2101), S6K (#2708), p-S6K (#9234), p38 (#9212), p-p38 (#9211) and cleavaged caspase 3 (#9661, Cell Signaling, Danver, MA, USA). The signal densities on the blots were measured with Image J (Wayne Rasband) and normalized using anti-α-tubulin antibody.
Hwang W.C., Kang D.W., Kang Y., Jang Y., Kim J.A, & Min D.S. (2020). Inhibition of phospholipase D2 augments histone deacetylase inhibitor-induced cell death in breast cancer cells. Biological Research, 53, 34.
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7

Comprehensive Western Blotting Antibody Panel

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Primary antibodies used for Western blotting were as follows: p-Tyr (sc-508), GAPDH (sc-32233), SHP2 (sc-7384), MCL-1 (sc-819) from Santa Cruz Biotechnology; p-EGFR (1068) (3777), p-ErbB2 (1248) (2247), ErbB3 (4754), p-ErbB3 (1289) (4791), p-ErbB3 (1328) (14525), p-ErbB4 (1284) (4757), p-Akt (308) (4056), p-Akt (473) (4060), p-Erk (202/204) (4370), p-P70S6K (389) (9205), p-4EBP1 (37/46) (2855), BIM (2933), BCL-xL (2764), p-P90RSK (380) (9341), p-S6 (235/6) (4858), p-S6 (240/4) (5364), c-Myc (5605), p-SHP2 (542) (3751), GAB1 (3232), p-GAB1 (627) (3233), p-GAB1 (659) (12745), β-ACTIN (4970), from Cell Signaling Technology. Secondary antibodies used were mouse IgG (GE Healthcare Life Sciences; NXA931) and rabbit IgG (GE Healthcare Life Sciences; NA934). Alpelisib and trametinib were purchased from AbMole Biosciences. RMC-4550 was purchased from Selleckchem. The SHP2 inhibitor (SHP099) for cell culture and in vivo experiments was purchased from MedchemExpress and AbMole Biosciences and was dissolved in DMSO at a stock concentration of 10 mmol/L for in vitro experiments. Human recombinant growth factors epiregulin (EREG) was purchased from Sigma-Aldrich (SRP3033) and neuregulin-1 (5898-NR) was purchased from R&D Systems.
Kurupi R., Floros K.V., Jacob S., Chawla A.T., Cai J., Hu B., Puchalapalli M., Coon C.M., Khatri R., Crowther G.S., Egan R.K., Murchie E., Greninger P., Dalton K.M., Ghotra M.S., Boikos S.A., Koblinski J.E., Harada H., Sun Y., Morgan I.M., Basu D., Dozmorov M.G., Benes C.H, & Faber A.C. (2022). Pharmacologic Inhibition of SHP2 Blocks Both PI3K and MEK Signaling in Low-epiregulin HNSCC via GAB1. Cancer Research Communications, 2(9), 1061-1074.
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8

Western Blot Analysis of Protein Expression

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Western blot analysis was performed as previously described.55 (link) Briefly, whole-cell lysate was prepared in the lysis buffer (RIPA), protein was quantified and equal amount (20–60 ug) of total protein was resolved by 10% SDS-polyacrylamide gel electrophoresis and/or 2% horizontal SDS agarose (for mucin) electrophoresis. Proteins were transferred to polyvinylidene difluoride membrane and incubated overnight with the primary antibody after blocking with 5% skim milk. Next day, blot was washed three times with PBST (0.1% Tween 20; (phosphate-buffered saline (PBS) with Tween 20)) and incubated with horseraddish peroxidase-conjugated secondary antibody for 1 h at room temperature. The proteins were visualized by enhanced chemiluminescent substrate (Thermo Scientific, Rockford, IL, USA). The primary antibodies used were NCOA3 (sc25742, Santa Cruz Biotechnologies, Dallas, TX, USA) PGK (sc-17943, Santa Cruz Biotechnologies), PARP (sc8007, Santa Cruz Biotechnologies), MUC4 (8G7, developed in our laboratory previously), EGFR (sc-03, Santa Cruz Biotechnologies) p-EGFR (S1046, sc-101665, Santa Cruz Biotechnologies), p-EGFR (Y1068, 3777, Cell Signaling Technologies, Danvers, MA, USA), Her-2 (2165, Cell Signaling Technology) and MUC1 (HMFG2, kindly gifted by Dr Hollingsworth).
Kumar S., Das S., Rachagani S., Kaur S., Joshi S., Johansson S., Ponnusamy M., Jain M, & Batra S. (2014). NCOA3-mediated upregulation of mucin expression via transcriptional and post-translational changes during the development of pancreatic cancer. Oncogene, 34(37), 4879-4889.
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9

Signaling Pathways Analysis in Cell Lines

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JWH-015 and SR144528 were purchased from Tocris Bioscience. Antibodies used were P-AKT, E-cadherin (Cell Signaling), P-ERK, ERK, AKT, GAPDH, P-EGFR, EGFR, VCAM-1, STAT3 (Santa Cruz), N-cadherin, P-FAK (Abcam), P-STAT3 (BD Biosciences).
Ravi J., Wani N.A., Elbaz M., Nasser M.W, & Ganju R.K. (2016). CANNABINOID RECEPTOR-2 AGONIST INHIBITS MACROPHAGE INDUCED EMT IN NON-SMALL CELL LUNG CANCER BY DOWNREGULATION OF EGFR PATHWAY. Molecular carcinogenesis, 55(12), 2063-2076.
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10

Colorectal Cancer Cell Lines and Drug Assays

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Rhein was purchased from TagerMol (Shanghai, China) and its purity is 98.7%. DMSO was used as a solvent to dissolve all drugs. Human colon cancer cell lines (HCT116, SW620, RKO and DLD-1) were obtained from cell resources center of the Shanghai Institutes for Biological Sciences (Chinese Academy of Sciences, Shanghai, China). HCT116 and RKO cells were grown in McCoy’s 5A medium (Gibco/BRL; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco; Life Technologies, Carlsbad, California, USA) and 1% of antibiotic solution (100 units/mL penicillin and 100 μg/mL streptomycin) in a humidified atmosphere of 5% CO2 at 37°C. SW620 and DLD-1 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Fisher Scientific; Suzhou, China) with 10% FBS (Gibco; Life Technologies, Carlsbad, California, USA), 1% of antibiotic solution and incubated at 37°C with 5% CO2. The antibodies against P-STAT3, STAT3, P-EGFR, EGFR, CDC2, CyclinB1, CyclinD1, BCL-2, BAX, GAPDH, horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG and HRP-conjugated goat anti-mouse IgG were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).
Zhuang Y., Bai Y., Hu Y., Guo Y., Xu L., Hu W., Yang L., Zhao C., Li X, & Zhao H. (2019). Rhein sensitizes human colorectal cancer cells to EGFR inhibitors by inhibiting STAT3 pathway. OncoTargets and therapy, 12, 5281-5291.
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