Journal: European journal of immunology

Article Title: Endoglycosidase treatment abrogates IgG arthritogenicity: importance of IgG glycosylation in arthritis.

doi: 10.1002/eji.200737581

Figure Lengend Snippet: Figure 1. Analysis of EndoS-treated CII-specific mAb. SDS-PAGE and lectin blot analysis of IgG mAb (M2139 – IgG2b and CIIC1 – IgG2a) incubated with and without EndoS and separated by 10% SDS-PAGE. The proteins were detected by Coomassie Brilliant Blue staining (Stain) or by blotting onto a membrane that was probed with LCA lectin (Blot).

Article Snippet: The membranes were blocked with 10 mM HEPES (pH 7.5) with 0.15 M NaCl, 0.01 mM MnCl2, 0.1 mM CaCl2, and 0.1% Tween-20 (HBST) and incubated with 1 lg/mL biotinylated LCA lectin (Vector Laboratories, Burlingame, CA).

Techniques: SDS Page, Incubation, Staining, Membrane

Journal: European journal of immunology

Article Title: Endoglycosidase treatment abrogates IgG arthritogenicity: importance of IgG glycosylation in arthritis.

doi: 10.1002/eji.200737581

Figure Lengend Snippet: Figure 2. EndoS-treated Ab bind to cartilage in vivo. Neonatal rats 1–2 days old were injected with 1 mg of CII-binding Ab [both normal and EndoS-treated (D)]) i.p. At 24 h after the Ab transfer, paws were dissected and snap-frozen in OCT compound using isopentane and dry ice. Immunohistochem- ical analysis was performed using biotinylated anti-mouse kappa (187.1) Ab and streptavidin peroxidase as detecting system using standard protocols. Stained joint sections (10 lm) from neonatal rats injected with M2139 (A), M2139D (B), CIIC1 (C), CIIC1D (D) and control, untreated rats (E) are shown. Magnification 10.

Article Snippet: The membranes were blocked with 10 mM HEPES (pH 7.5) with 0.15 M NaCl, 0.01 mM MnCl2, 0.1 mM CaCl2, and 0.1% Tween-20 (HBST) and incubated with 1 lg/mL biotinylated LCA lectin (Vector Laboratories, Burlingame, CA).

Techniques: In Vivo, Injection, Binding Assay, Staining, Control