Human RGM-A Antibody

RGM‑A in Human Brain.

RGM-A was detected in immersion fixed paraffin-embedded sections of human brain (spinal cord) using Goat Anti-Human RGM-A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2459) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). 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 the dorsal horn. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of RGM-A by Western Blot

HFE2 administration significantly improves EAE disease outcome. A Western Blot analysis shows a strong HFE2 reduction in EAE animal serum when compared to control. ELISA quantification showing HFE2 decrease and RGMa increase in EAE animal serum (mean ± s.d.; EAE n = 3 and RGMa n = 3). b Body conditioning assessment (‘0’ no paralysis to ‘4’ forelimb weakness) of EAE-induced mice with HFE2 + /- RGMa treatment every 3 days (d3, 6, 9, 12 and 15; mean ± s.e.m.; unpaired two-tail t-tests; PBS n = 8, HFE2 n = 6, RGMa+ HFE2 n = 8). c Body conditioning assessment of EAE-induced Hfe2fl/fl and Hfe2 deltaAlb-cre mice (mean ± s.e.m.; unpaired two-tail t-tests; Hfe2fl/fln = 8 and Hfe2 deltaAlb-cren = 8). d Representative confocal images of CD3 + , B220+ and CD11b+ immune staining in spinal cord of EAE-induced mice 3 weeks after induction and quantification (mean ± s.e.m.; unpaired two-tail t-test; PBS n = 7, HFE2 n = 6 mice; technical replicates shown). e Schematic of activated immune cell adoptive transfer. Representative confocal images of CD4-positive labelled transferred cell (CellVue) in PBS-treated or HFE2-treated recipient WT mice and quantification (mean ± s.e.m.; unpaired two-tail t-test; PBS n = 6, HFE2 n = 6). Ptx; Pertussis Toxin. f Representative confocal images of fibrinogen in the spinal cord of HFE2-treated EAE-mice and quantification (mean ± s.e.m.; unpaired two-tail t-test; PBS-treated n = 6 and HFE2 n = 6). *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file (includes exact p-values). Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-024-45303-1), licensed under a CC-BY license. Not internally tested by R&D Systems.