Journal:

Article Title: Immunoglobulin A Antibodies against Ricin A and B Subunits Protect Epithelial Cells from Ricin Intoxication

doi: 10.1128/IAI.02088-05

Figure Lengend Snippet: Anti-RTB IgA MAbs block ricin attachment to ASF. Biotinylated ricin (8 ng/ml) was mixed with each of the indicated MAbs at a saturating concentration (10 μg/ml) and overlaid in triplicate onto ELISA plates coated with ASF. The plates were then washed and developed using avidin-HRP and TMB, as described in Materials and Methods. One hundred percent binding was defined as the value obtained for wells probed with ricin in the absence of antibody. Each bar represents the average, with SE, of two samples performed in duplicate.

Article Snippet: Unlabeled and biotinylated derivatives of ricin ( Ricinus communis agglutinin II), as well as polyclonal goat anti-RCA-I/II antiserum, were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Blocking Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Avidin-Biotin Assay, Binding Assay

Journal:

Article Title: Immunoglobulin A Antibodies against Ricin A and B Subunits Protect Epithelial Cells from Ricin Intoxication

doi: 10.1128/IAI.02088-05

Figure Lengend Snippet: Anti-RTB IgA MAbs inhibit ricin binding to the luminal surfaces of the human duodenum. Biotinylated ricin (5 μg/ml) was incubated with each of the indicated IgA MAbs (20 μg/ml) and then overlaid for 1 h onto deparaffinized tissue sections of human duodenum. The sections were washed, labeled with streptavidin-FITC, and visualized by fluorescence microscopy. (A) Ricin uniformly labeled the luminal surfaces of human intestinal villi. Occasional goblet cells were also labeled (bottom center). (B) 33G2 reduced ricin binding to luminal surfaces to undetectable levels. (C) 35H6 reduced toxin binding to luminal surfaces, but residual staining was observed on the surrounding glass surfaces. Neither 23D7 (D) nor TEPC-15 (E) had an effect on ricin attachment to tissue sections compared to sections treated with ricin only. (F) Tissue section not labeled with ricin. Bar, 20 μm.

Article Snippet: Unlabeled and biotinylated derivatives of ricin ( Ricinus communis agglutinin II), as well as polyclonal goat anti-RCA-I/II antiserum, were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Binding Assay, Incubation, Labeling, Fluorescence, Microscopy, Staining

Journal:

Article Title: Immunoglobulin A Antibodies against Ricin A and B Subunits Protect Epithelial Cells from Ricin Intoxication

doi: 10.1128/IAI.02088-05

Figure Lengend Snippet: Anti-RTA IgA MAbs reduce transcytosis of ricin across polarized epithelial cell monolayers. Biotinylated ricin (200 ng) was incubated with the indicated IgA MAbs (2 μg in DMEM) in a volume of 0.2 ml for 1 h and then applied to the apical surfaces of polarized MDCK II cell monolayers. Eighteen hours later, ricin in the basolateral compartments was collected using avidin-agarose beads, which were then subjected to SDS-PAGE and Western blot analysis, as described in Materials and Methods. As positive and negative controls, samples of DMEM containing ricin (200 ng) (lane labeled “standard”) or no ricin (negative control) were treated in parallel.

Article Snippet: Unlabeled and biotinylated derivatives of ricin ( Ricinus communis agglutinin II), as well as polyclonal goat anti-RCA-I/II antiserum, were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Incubation, Avidin-Biotin Assay, SDS Page, Western Blot, Labeling, Negative Control