Anti egfr

Manufactured by R&D Systems

Sourced in United States

Anti-EGFR is a laboratory reagent used in research applications. It is an antibody that binds to the Epidermal Growth Factor Receptor (EGFR) protein. Anti-EGFR can be used to detect and quantify EGFR expression in various cell and tissue samples.

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6 protocols using anti egfr

Protein Expression Analysis by Western Blot

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Protein extracts were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred to polyvinylidene fluoride membranes (Millipore, Germany). Protein expression was detected by western blot analysis. Antibodies used herein including anti-IL-6, anti-TNF-α, anti-p-AKT, anti-AKT, anti-SRC, anti-p-SRC, anti-VEGFA, anti-MAPK1, anti-IL-1β, anti-EGFR and β-actin were obtained from R&D Systems. Band density was quantified using ImageJ software and normalised to the corresponding control group.

Wang Y., Gu W., Kui F., Gao F., Niu Y., Li W., Zhang Y., Guo Z, & Du G. (2021). The mechanism and active compounds of semen armeniacae amarum treating coronavirus disease 2019 based on network pharmacology and molecular docking. Food & Nutrition Research, 65, 10.29219/fnr.v65.5623.

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Western Blot Analysis of Signaling Proteins

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The primary antibodies anti‐ALK (C26G7), anti‐phospho‐ALK (Tyr1604), anti‐phospho‐HER3 (Tyr1289), anti‐HER3, anti‐phospho‐Akt (S473), anti‐Akt, anti‐phospho‐epidermal growth factor receptor (EGFR), anti‐phospho‐MET (Tyr1234/1235), anti‐MET, and anti‐β‐actin were obtained from Cell Signaling Technology. Anti‐ERK1/ERK2, p‐ERK1/ERK2 (T202/Y204), and anti‐EGFR were obtained from R&D Systems. Western blot analysis was performed as described previously.²⁰

Taniguchi H., Akagi K., Dotsu Y., Yamada T., Ono S., Imamura E., Gyotoku H., Takemoto S., Yamaguchi H., Sen T., Yano S, & Mukae H. (2022). Pan‐HER inhibitors overcome lorlatinib resistance caused by NRG1/HER3 activation in ALK ‐rearranged lung cancer. Cancer Science, 114(1), 164-173.

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Single-Cell Multiomics Assay Protocol

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HCT116, A549, H1975, H1650, and HCC827 cell lines were purchased from the cell bank of Chinese Academy of Sciences. HCT116 cells were routinely maintained in ATCC-formulated McCoy’s 5A modified medium containing 10% (v/v) fetal bovine serum (FBS) in humidified atmosphere of 5% CO2 and 95% air at 37 °C. A549, H1650, H1975, and HCC827 cells were routinely maintained in RPMI 1640 medium supplemented with 10% (v/v) FBS. Size- and color-encoded Fluoresbrite Carboxylate beads (see Table S1) and PolyLink Protein Coupling Kit were purchased from Polyscience, Inc. Antibody pairs used in single-cell assays were listed in Table S2. Alexa Fluor 647 Antibody Labeling Kit was purchased from Life Technologies. Biotinylated antibodies (anti-EpCAM and anti-EGFR) were purchased from R&D Systems. Streptavidin-coated magnetic beads (0.8 μm) were purchased from Solulink. The 0.1% (w/v) PLL solution was purchased from Sigma-Aldrich. PLL-coated glass slides were obtained from Thermo Fisher Scientific. Three sets of optical filter modules (365 nm/410 nm, 440 nm/485 nm, 530 nm/565 nm) were purchased from Chroma Technology Corp.

Yang L., Wang Z., Deng Y., Li Y., Wei W, & Shi Q. (2016). Single-Cell, Multiplexed Protein Detection of Rare Tumor Cells Based on a Beads-on-Barcode Antibody Microarray. Analytical chemistry, 88(22), 11077-11083.

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Comprehensive Protein Expression Analysis

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The primary Abs, including anti‐EGFR, anti‐MET, anti‐phospho‐MET, anti‐HER2, anti‐phospho‐HER2, anti‐phospho‐Akt (S473), anti‐Akt, anti‐E‐cadherin, anti‐Vimentin, anti‐ZEB1, anti‐Snail, anti‐Slug, anti‐GSK3α/β, anti‐phospho‐GSK3α/β (ser21/ser9), anti‐β‐catenin, anti‐cleaved caspase3, and anti‐β‐actin were obtained from Cell Signaling Technology. The anti‐ERK1/ERK2, anti‐p‐ERK1/ERK2 (T202/Y204), and anti‐EGFR Abs were obtained from R&D Systems. Western blot analysis was carried out as previously described.¹⁰

Fukuda K., Takeuchi S., Arai S., Kita K., Tanimoto A., Nishiyama A, & Yano S. (2020). Glycogen synthase kinase‐3 inhibition overcomes epithelial‐mesenchymal transition‐associated resistance to osimertinib in EGFR‐mutant lung cancer. Cancer Science, 111(7), 2374-2384.

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Functionalization of Gadolinium Boride Nanoparticles

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In a typical experiment, 3 mg of Gd¹⁰B₆NPs and 20 mg of F127- COOH were added to the solution at
room temperature containing 2 mL of chloroform and evaporated under
vacuum. In the next step, 5 mg of F127-COOH-capped Gd¹⁰B₆NPs was mixed with 1-ethyl-3-(3-(dimethylamino)propyl)
carbodiimide (EDC) (3 mg, 15.8 μmol) and N-hydroxy
succinimide (NHS) (2 mg, 17.4 μmol) in 1× PBS solution.
To this mixture was added 20 μL (50 μg/mL) of anti-EGFR
(R&D Systems, Minneapolis, USA), and the mixture was stirred for
4 h at 4 °C. The obtained anti-EGFR-functionalized Gd¹⁰B₆NPs were washed thoroughly to remove the unreacted anti-EGFR
and then redispersed and stored in 1× PBS for further use.

Shanmugam M., Kuthala N., Kong X., Chiang C.S, & Hwang K.C. (2023). Combined Gadolinium and Boron Neutron Capture Therapies for Eradication of Head-and-Neck Tumor Using Gd10B6 Nanoparticles under MRI/CT Image Guidance. JACS Au, 3(8), 2192-2205.

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Enrichment and Detection of Extracellular Vesicles

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Pyrrole, poly(sodium 4-styrenesulfonate) (PSS), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), streptavidin, and sodium hydroxide (NaOH) were obtained from Sigma Aldrich (St. Louis, MO, USA). Anodized aluminum oxide (AAO) membrane filter (pore diameter, 200 nm) was purchased from Whatman (Pittsburgh, PA, USA). Silver paste (P-100 type) was obtained from CANS. NHS-SS-biotin was purchased from CovaChem (Loves Park, Illinois, USA). Biotinylated anti-EpCAM, anti-EGFR, anti-vimentin, anti-N-cadherin, and anti-Trop2 were obtained from R&D system (Minneapolis, MN, USA). Biotinylated anti-CD63 and anti-CD81 were purchased from AnCell (Oak Park, Minnesota, USA). Biotinylated anti-CD9 was obtained from Abcam (Cambridge, UK).

Lim J., Choi M., Lee H., Han J.Y, & Cho Y. (2019). A Novel Multifunctional Nanowire Platform for Highly Efficient Isolation and Analysis of Circulating Tumor-Specific Markers. Frontiers in Chemistry, 6, 664.

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