Mouse Osteoprotegerin/TNFRSF11B Antibody

Osteoprotegerin/TNFRSF11B Inhibition of TRAIL/TNFSF10-induced Cytotoxicity and Neutralization by Mouse Osteoprotegerin/TNFRSF11B Antibody. In the presence of the metabolic inhibitor actinomycin D (1 µg/mL), Recombinant Mouse OPG/TNFRSF11B Fc Chimera (Catalog # 459-MO) inhibits Recombinant Human TRAIL/TNFSF10 (Catalog # 375-TL) induced cytotoxicity in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Under these conditions, inhibition of Recombinant Human TRAIL/TNFSF10 (20 ng/mL) activity elicited by Recombinant Mouse OPG/TNFRSF11B Fc Chimera (0.1 µg/mL) is neutralized (green line) by increasing concentrations of Mouse OPG/TNFRSF11B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF459). The ND₅₀ is typically 0.15-0.9 µg/mL.

Osteoprotegerin/TNFRSF11B in Mouse Stomach. Osteoprotegerin/TNFRSF11B was detected in perfusion fixed frozen sections of mouse stomach using Goat Anti-Mouse Osteoprotegerin/TNFRSF11B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF459) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to glandular epithelial cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse Osteoprotegerin/TNFRSF11B by Western Blot Cell-autonomous defect of Col1a1-Krm2 transgenic osteoblasts.(A) BrdU incorporation assays revealed a higher proliferation rate in primary calvarial osteoblast cultures from transgenic mice after 2 days of differentiation. (B) Von Kossa staining performed at 10 days of differentiation reveals reduced mineralization of osteoblasts from Col1a1-Krm2 mice (scale bars, 1 cm), despite higher protein content (given below). Values represent mean ± SD (n = 3). Asterisks indicate statistically significant differences. (C) Western Blot analysis of canonical Wnt signaling using primary osteoblasts from wildtype and transgenic mice following stimulation with Wnt3a for 30 minutes. (D) Western Blot analysis (left) and ELISA (right) showing decreased Opg levels in cellular extracts and conditioned medium of osteoblasts from Col1a1-Krm2 mice. (E) Quantitative RT-PCR (left) and ELISA (right) demonstrating that the reduced expression of Tnfrsf11b in bones of 6 weeks old Col1a1-Krm2 mice results in decreased Opg serum levels. All bars represent mean ± SD (n = 4). Asterisks indicate statistically significant differences. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20436912), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Osteoprotegerin/TNFRSF11B by Western Blot Cell-autonomous defect of Col1a1-Krm2 transgenic osteoblasts.(A) BrdU incorporation assays revealed a higher proliferation rate in primary calvarial osteoblast cultures from transgenic mice after 2 days of differentiation. (B) Von Kossa staining performed at 10 days of differentiation reveals reduced mineralization of osteoblasts from Col1a1-Krm2 mice (scale bars, 1 cm), despite higher protein content (given below). Values represent mean ± SD (n = 3). Asterisks indicate statistically significant differences. (C) Western Blot analysis of canonical Wnt signaling using primary osteoblasts from wildtype and transgenic mice following stimulation with Wnt3a for 30 minutes. (D) Western Blot analysis (left) and ELISA (right) showing decreased Opg levels in cellular extracts and conditioned medium of osteoblasts from Col1a1-Krm2 mice. (E) Quantitative RT-PCR (left) and ELISA (right) demonstrating that the reduced expression of Tnfrsf11b in bones of 6 weeks old Col1a1-Krm2 mice results in decreased Opg serum levels. All bars represent mean ± SD (n = 4). Asterisks indicate statistically significant differences. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20436912), licensed under a CC-BY license. Not internally tested by R&D Systems.