Human Epiregulin Antibody

Cell Proliferation Induced by Epiregulin and Neutralization by Human Epiregulin Antibody. Recombinant Human Epiregulin (Catalog # 1195-EP) stimulates proliferation in the Balb/3T3 mouse embryonic fibroblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Epiregulin is neutralized (green line) by increasing concentrations of Goat Anti-Human Epiregulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1195). The ND₅₀ is typically ≤1.5 µg/mL.

Human Epiregulin ELISA Standard Curve. Recombinant Human Epiregulin protein was serially diluted 2-fold and captured by Rat Anti-Human Epiregulin Monoclonal Antibody (Catalog # MAB14252) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Epiregulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1195) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Detection of Human Epiregulin by Immunohistochemistry EREG expression correlates with MMP-1 levels in human DCIS lesions. a qRT-PCR analysis of MMP-1 gene expression in MCF10A and MCF10DCIS cells. Expression levels of MMP-1 are normalized to CYBP. b Immunoblot analysis of MMP-1 in conditioned media obtained from MCF10A and MCF10DCIS cells. Loading was assessed by Coomassie staining (Additional file 1: Figure S1C). c qRT-PCR analysis of MMP-1 gene expression in MCF10DCIS cells expressing either NT or EREG shRNA constructs. Expression levels of MMP-1 are normalized to CYBP. d Immunohistochemistry of normal breast tissue and DCIS stained with an anti-EREG antibody. Scale bars represent 50 μm. e Normal and DCIS human samples were stained with an anti-MMP-1 antibody. Representative images of normal and varying levels of MMP-1 staining in DCIS lesions are shown. f Quantification of the percent of samples staining positive for EREG. g Quantification of the percent of samples staining positive for MMP-1. Scale bars represent 50 μm. *p < 0.05. ***p < 0.0001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26215578), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Epiregulin by Immunocytochemistry/ Immunofluorescence EREG increased IL-8 production from cancer cells through EGFR signaling to induce tumor neovascularization. (A) Representative images of the change in angiogenic factors in both Huh7 and HepG2 cells with/without stimulation with EREG. IL-8 levels in Huh7 cells only increased during stimulation with rEREG. Red box showed the expression of IL-8. The graph for the intensity of IL-8 in the angiogenesis array kit in the untreated and EREG-treated groups (B) in Huh7 and (C) in HepG2. The graph for the expression level of IL-8 by RT-PCR in the untreated and EREG-treated groups (D) in Huh7 and (E) in HepG2. (F) The expression level of IL-8 in Huh7 cells in the monoculture group (left graph) and the group co-cultured with LX-2 (right graph) under the LPS stimulation. (G) Change in IL-8 expression levels when treated with/without anti-EREG antibodies under LPS present. (H) Representative images of the change in angiogenic factors in LX-2 with/without stimulation with LPS. (I) The graph for the intensity of IL-8 measured by the angiogenesis array kit in the untreated and LPS-treated groups in LX-2. Significance was determined using a one-way analysis of variance for group comparisons. * p < 0.05, ** p < 0.01, **** p < 0.0001, ns; not significant. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/25/8/4405), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Epiregulin by Western Blot EREG promoted cell proliferation and migration/invasion activity of liver cancer cells expressing EGFR in vitro. (A) Expression levels of EGFR in each HCC line (Huh7, JHH, and HepG2). EGFR was expressed in Huh7 and JHH cells but not in HepG2 cells. Cell proliferation of (B) Huh7 cells and (C) HepG2 cells was stimulated with different concentrations of EREG (0, 1, 10, 50, and 100 ng/mL). (D) Cell proliferation of Huh7 co-cultured with LX-2 was measured by the WST-1 assay when stimulated with 100 and 1000 ng/mL of LPS. (E) Change in the phosphorylation of ERK1/2 in Huh7 stimulated with EREG and in combination with EREG and EGFR inhibitors (AG1478; 100 ng/mL). (F) Cell proliferation of Huh7 co-cultured with LX-2 with LPS stimulation and a combination of LPS and anti-EREG antibodies. Significance was determined using a one-way analysis of variance for group comparisons. *; p < 0.05, **; p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/25/8/4405), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Epiregulin by Immunocytochemistry/ Immunofluorescence Change in EREG expression and production in LX-2 cells by LPS stimulation. (A) Expression level of EREG in LX-2 stimulated with different concentrations of LPS (0, 1, 10, and 100 ng/mL) measured by RT-PCR. (B) Expression of EREG in LX-2 cells measured by RT-PCR and (C) production of EREG protein level by enzyme-linked immuno-sorbent assay (ELISA) in the supernatant with LX-2 cells cultured with/without 100 ng/mL of LPS for 24 h. (D) EREG expression in LX-2 cells stimulated with/without recombinant human EREG protein (100 ng/mL) by RT-PCR. (E) Representative images of immunostaining for EREG in LX-2 cells compared between the conditions stimulated with/without LPS stimulation. Magnification, 60×. (F) Gene expression levels of other members in the EGF family in LX-2 cells with/without LPS stimulation. Graphs showed the mean ± SEM. *; p < 0.05, ***; p < 0.001, ns; not significant. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/25/8/4405), licensed under a CC-BY license. Not internally tested by R&D Systems.