Human RANK/TNFRSF11A Antibody

Human RANK/TNFRSF11A Antibody Agonist Activity

Human RANK/TNFRSF11A Antibody (Catalog # AF683) induces osteoclast differentiation of RAW 264.7 mouse monocyte/macrophage cells in the presence of recombinant mouse M-CSF (416ML). The ED₅₀ for this effect is typically 0.0750 – 1.50 μg/mL.

Detection of RANK/TNFRSF11A by Western Blot

Expression of mRNA encoding tumor necrosis factor receptor superfamily, member 11a (TNFRSF11A, wild type receptor activator of NF-kB, wt RANK), TNFRSF11A_ delta 9 (RANK-a), TNFRSF11A_ delta 8,9 (RANK-b) and TNFRSF11A_ delta 7,8,9 (RANK-c) in breast cancer cell lines and breast cancer samples. A. Agarose gel electrophoresis (2%) of PCR products using primers P1 and P2 depicting TNFRSF11A and the identified alternative splice variants in MDA-MB-231, SKBR3, MCF7, MDA-MB-468, T47D and MCF10A breast cancer cell lines. vTNFRSF11A_exon9a was observed only in SKBR3 and after cloning and sequencing was identified as a TNFRSF11A variant encompassing exons 6, 7, 8, 9a and 10. Splice variant TNFRSF11A_ delta 7,8,9 is in bold. B. TNFRSF11A_ delta 7,8,9 (RANK-c) relative mRNA expression levels in a panel of breast cancer cell lines and the non-tumorigenic epithelial cell line MCF10A; values obtained for MCF10A were set to 100. C. Relative mRNA expression levels of variant TNFRSF11A_ delta 7,8,9 (RANK-c) in a panel of formaldehyde-fixed paraffin embedded (FFPE) invasive ductal breast carcinoma samples analyzed against tumor histological grade and proliferation index Ki-67, respectively. D. Relative mRNA expression levels of TNFRSF11A (wt RANK) in the same panel of invasive ductal breast carcinoma samples plotted against tumor histological grade. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22824341), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human RANK/TNFRSF11A by Western Blot

Induction of the expression of RANK-L, OPG, and RANK by normal human blood neutrophils incubated in the presence of synovial fluid (SF) from patients with rheumatoid arthritis (RA) or patients with osteoarthritis (OA). (a) Surface expression of RANK-L by normal blood neutrophils incubated in SF from patients with RA (RA-SF) or OA (OA-SF) for 2 days. Flow cytometry was performed after incubation of neutrophils with a goat anti-human RANK-L antibody followed by a fluorescein isothiocyanate (FITC)-conjugated anti-goat F(ab')2 antibody. Control isotype antibody was a normal goat immunoglobulin G (IgG). Results shown are representative of three RA-SF and nine OA-SF. (b) Expression of mRNA for OPG and RANK by normal blood neutrophils incubated for 2 days in SF from four patients with RA and two patients with OA. Total RNA was isolated from freshly isolated normal blood neutrophils (D0) and from neutrophils of the same healthy donors after 2 days of incubation in SF (RA-1 to -4, OA-1, -2). RNA was then analyzed by reverse transcriptase-polymerase chain reaction. Results shown are representative of two different healthy donors. (c) Surface expression of RANK by normal blood neutrophils incubated in SF from patients with RA (RA-SF) or OA (OA-SF) for 3 days. Flow cytometry was performed after cellular fixation, permeabilization, and staining with a mouse monoclonal anti-human RANK antibody followed by a FITC-conjugated anti-mouse F(ab')2 antibody. Control isotype antibody was a non-specific mouse IgG. Results shown are representative of neutrophils from two different healthy subjects incubated in three different RA-SF and two OA-SF. OPG, osteoprotegerin; RANK, receptor activator of nuclear factor-kappa-B; RANK-L, ligand of receptor activator of nuclear factor-kappa-B. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/17341304), licensed under a CC-BY license. Not internally tested by R&D Systems.