Goat anti egfr

Manufactured by R&D Systems

Sourced in Germany

Goat anti-EGFR is a laboratory reagent that binds to the Epidermal Growth Factor Receptor (EGFR) protein. It is produced in goats and can be used in various immunodetection and analysis techniques.

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Anti-EGFR (6 citations)

4 protocols using goat anti egfr

Immunohistochemical Analysis of SVZ and Olfactory Bulb

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Brains were fixed overnight at 4°C in 4% PFA in PBS, then cryoprotected in 30% sucrose in PBS for 1–3 days at 4°C, then frozen in OCT or cryogel on dry ice after 3–12 hr equilibration at 4°C. Sections were cut at 12 μm thickness spanning the rostral half of the SVZ (typically 6 sections per slide) or the entire olfactory bulb (typically 8 sections per slide). Sections were immunostained with the following primary antibodies: rat anti-BrdU (Abcam clone Bu1/75, 1/500, after heat-mediated antigen retrieval), guinea pig anti-Dcx (1/1000; Millipore), mouse anti-Mcm2 (BD Biosciences, 1/500, after heat-mediated antigen retrieval), rat anti-Ki67 (1/500; eBioscience), mouse anti-tyrosine hydroxylase (1/1000; Millipore), rabbit anti-calretinin (1/1000; Sigma), rabbit anti-calbindin (1/500; Millipore), rabbit anti-S100β (1/1000; Dako), rabbit anti-GST-pi (1/3000; Enzo, Farmingdale, NY), and mouse anti-NeuN (1/1000; Millipore). Fixed whole-mount SVZs were stained with mouse anti-acetylated tubulin (1/1000; Sigma), rabbit anti-β-catenin (1/500; Sigma), mouse anti-GFAP (1/3000; Sigma), and goat anti-EGFR (1/250; R&D Systems). Alexa Fluor 488-, 555-, and 647-conjugated secondary antibodies were used (Life Technologies, Carlsbad, CA).

Mich J.K., Signer R.A., Nakada D., Pineda A., Burgess R.J., Vue T.Y., Johnson J.E, & Morrison S.J. (2014). Prospective identification of functionally distinct stem cells and neurosphere-initiating cells in adult mouse forebrain. eLife, 3, e02669.

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Multiplex Immunohistochemistry Protocol

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Fluorescence immunohistochemistry utilized a citrate-based antigen retrieval step involving pH 6.0 sodium citrate in water heated to 95 °C for 7 min, followed by cooling to room temperature and immunostained as described previously (38 (link)). Antibodies and dilutions are as follows: rabbit anti-Pax6 (Millipore; 1:750), rabbit Hopx (Sigma; 1:500); mouse anti-Ki67 (Vector; 1:300), goat anti-EGFR (R&D Systems; 10 mg/mL); rabbit anti-Olig2 (Abcam; 1:2,000), rabbit anti-Tbr1 (Abcam; 1:1,000), rat anti-GFAP (1:100), mouse anti-βIII-tubulin (R&D Systems; 1:1,000), chicken anti-Tbr2 (Millipore; 1:200), rat anti-BrdU (Accurate Chemical; 1:400); secondary antibodies and dilutions were donkey anti-goat/chicken/rat, 1:1,000, DyLight 488/543/647 (Jackson Laboratories). Blocking utilized reconstituted Donkey Serum. Sections were counterstained with DAPI mounting medium (Vector) to label cellular nuclei. A single injection of BrdU at 50 mg/kg (Sigma; in 0.9% NaCl with 0.007 N NaOH) was delivered intravenously at E97, with sacrifice 1 h later.

Rash B.G., Arellano J.I., Duque A, & Rakic P. (2022). Role of intracortical neuropil growth in the gyrification of the primate cerebral cortex. Proceedings of the National Academy of Sciences of the United States of America, 120(1), e2210967120.

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Quantitative EGFR Phosphorylation Assay

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Primary antibodies: Mouse monoclonal antibody PY72 (Glenney et al., 1988 (link)) (InVivo Biotech Services, Henningsdorf, Germany), rabbit anti EGFR pY₁₀₆₈ (Cell Signaling; 1:400), goat anti EGFR (R&D Systems; 1:300). Secondary antibodies: Alexa Fluor® 568 donkey anti-rabbit IgG (Life Technologies, 1:200), Alexa Fluor® 568 donkey anti-mouse IgG (Life Technologies, 1:200), Alexa Fluor® 647 donkey anti-goat IgG (Life Technologies, 1:200), Alexa Fluor® 647 chicken anti-mouse IgG (Life Technologies, 1:200), Alexa Fluor® 647 donkey anti-rabbit IgG (Life Technologies, 1:200), IRDye® 800CW Donkey anti-Rabbit IgG (Licor, 1:10000), IRDye® 680RD Donkey anti-Mouse IgG (Licor, 1:10000).

Stanoev A., Mhamane A., Schuermann K.C., Grecco H.E., Stallaert W., Baumdick M., Brüggemann Y., Joshi M.S., Roda-Navarro P., Fengler S., Stockert R., Roßmannek L., Luig J., Koseska A, & Bastiaens P.I. (2018). Interdependence between EGFR and Phosphatases Spatially Established by Vesicular Dynamics Generates a Growth Factor Sensing and Responding Network. Cell Systems, 7(3), 295-309.e11.

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Antibody Reagents for Cellular Signaling

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rabbit anti‐RPTPγ (rb‐RPTPγ‐P4: gift from C. Sorio, Department of Pathology, University of Verona, Verona, Italy, (Sorio et al, 1995 (link))), mouse anti‐TCPTP (MAB1930, R&D Systems, Minneapolis, MN), mouse anti‐p22^phox (ab80896, Abcam, Cambridge, UK), anti‐cysteine sulfenic acid 2‐thiodimedone (ABS30, Merck, Darmstadt, Germany), living colors mouse anti‐GFP (632381, Clontech, Mountain View, CA), goat anti‐GFP (ab5450, Abcam), rabbit anti‐phospho EGFR Y1068 (3777, Cell Signaling Technology, Danvers, MA), mouse anti‐phospho EGFR Y1068 (2236, Cell Signaling Technology) rabbit anti‐EGFR (4267, Cell Signaling Technology), goat anti‐EGFR (AF231, R&D Systems), rabbit anti‐phospho‐ERK‐1/2 Thr/Tyr 202/204 (9101, Cell Signaling Technology), mouse anti‐ERK1/2 (ab366991, Abcam), rabbit anti‐phospho‐Akt Ser473 (9271, Cell Signaling Technology), mouse anti‐Akt (pan) (2920, Cell Signaling Technology), rabbit anti‐EEA1 (3288, Cell Signaling Technology), rabbit anti‐Rab7 (9367, Cell Signaling Technology), rabbit anti‐Rab11a (2413, Cell Signaling Technology), rabbit phospho‐Rb Ser807/811 (8516, Cell Signaling Technology), mouse anti‐GAPDH (CB1001, Merck), mouse anti‐α‐Tubulin (T6074, Merck), mouse anti‐Na⁺/K⁺ ATPase‐α3‐subunit (BML‐SA247‐0100, Enzo Life Sciences, NY).

Joshi M.S., Stanoev A., Huebinger J., Soetje B., Zorina V., Roßmannek L., Michel K., Müller S.A, & Bastiaens P.I. (2023). The EGFR phosphatase RPTPγ is a redox‐regulated suppressor of promigratory signaling. The EMBO Journal, 42(10), e111806.

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