Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Recombinant human IgA1 and IgA2 autoantibodies to type VII collagen induce subepidermal blistering ex vivo.

doi: 10.4049/jimmunol.1400160

Figure Lengend Snippet: FIGURE 1. Recombinant autoantibodies demonstrated the correct m.w. under reducing and nonreducing conditions, labeled the DEJ and bound to type VII collagen extracted from human dermis. (A) Recombinant autoantibodies were separated by 12% SDS-PAGE under reducing conditions and stained with Coomassie blue. The H chains of the recombinant proteins migrated at 50 (rIgG1), 60 (rIgA1), and 55 kDa (rIgA2), and the k L chains migrated at 25 kDa. The m.w. standard is indicated on the left. (B) Equal amounts of recombinant autoantibodies were separated by 7% SDS-PAGE under nonreducing conditions. The majority of the recombinant autoantibodies were assembled as complete 2H2L heterotetramers. In addition, for rIgA1 and rIgA2, additional bands at higher m.w. were detectable, indicating polymerization and a reduced amount of monomeric rIgA1 or rIgA2. (C) Immunoblotting of recombinant autoantibodies was performed using the same conditions as in (B), as well as detection with peroxidase-conjugated polyclonal anti-human k L chain Ab. The Coomassie staining of rIgA1 and rIgA2 appeared to be weaker overall than that for rIgG1, which may be because of the higher glycosylation content of IgA isotypes. The m.w. standard is indicated on the left. (D) IgA1 and IgA2 recombinant autoantibodies reacted with a 290-kDa band after immunoblotting of dermal extracts, whereas the isotype controls for IgA1 (Iso IgA1) and IgA2 (Iso IgA2) were negative. (E) Human neonatal foreskin cryosections were incubated with the different recombinant autoantibodies and visualized by isotype- and subclass-specific FITC-conjugated Abs. Both rIgA1 and rIgA2 presented a defined reactivity with the DEJ (arrows). As controls, a V-gene matched rIgG1 (31) and serum from a healthy volunteer (NHS) in conjunction with FITC-conjugated polyclonal rabbit anti-human IgG was used. Original magnification 3100.

Article Snippet: For the detection of Abs in cryosections, FITC-conjugated polyclonal goat anti-human IgA (Bio-Rad Laboratories GmbH, Munich, Germany) and polyclonal goat anti-human IgG (Sigma-Aldrich Chemie GmbH, Munich, Germany) were used as secondary Abs.

Techniques: Recombinant, Labeling, SDS Page, Staining, Western Blot, Glycoproteomics, Incubation

Journal: Applied Sciences

Article Title: Association between Non-Alcoholic Steatohepatitis-Related Hepatocellular Carcinoma and Periodontopathic Bacteria: A Cross-Sectional Pilot Study

doi: 10.3390/app13063893

Figure Lengend Snippet: Figure 1. Salivary IgA levels: (a) saliva flow rate; (b) salivary IgA concentration; (c) salivary IgA flow rate. The presented values are medians (first quartile–third quartile), and the Mann–Whitney’s U test was used for statistical analysis (* p < 0.05). Each 99% confidence intervals were 0.972–0.980: (a), 0.005–0.010: (b) and 0.001–0.004: (c).

Article Snippet: Salivary IgA concentration was measured by ELISA using the Human IgA ELISA kit (Bethyl Laboratories, Montgomery, AL, USA) following the manufacturer’s protocol.

Techniques: Concentration Assay

Journal: Cell Reports Medicine

Article Title: The Fc-effector function of COVID-19 convalescent plasma contributes to SARS-CoV-2 treatment efficacy in mice

doi: 10.1016/j.xcrm.2022.100893

Figure Lengend Snippet:

Article Snippet: Peroxidase AffiniPure Goat Anti-Human IgA + IgG + IgM (H + L) , Jackson ImmunoResearch , 109-035-064.

Techniques: Blocking Assay, Control, Virus, Variant Assay, Recombinant, Modification, Saline, Red Blood Cell Lysis, Lysis, Electron Microscopy, Staining, Luciferase, Multiplex Assay, cDNA Synthesis, Oligo Synthesis, Plasmid Preparation, Expressing, Software, Flow Cytometry, Real-time Polymerase Chain Reaction, Spectrophotometry, Membrane, Stripping Membranes, Isolation, In Vivo Imaging