Journal: PLoS ONE
Article Title: Natural Pig Plasma Immunoglobulins Have Anti-Bacterial Effects: Potential for Use as Feed Supplement for Treatment of Intestinal Infections in Pigs
Figure Lengend Snippet: Characterisation of ppIgG. (A) Pig plasma IgG product (ppIgG, 1 mg/ml) was run under reducing conditions on 12% SDS PAGE as described in Materials and Methods. In parallel, Western blotting (WB) was performed on the same sample. The blot was developed with biotinylated rabbit anti-pig IgG F(ab) 2 antibody, followed by alkaline phosphatase-coupled streptavidin (see Materials and Methods ). (B) Wells were coated with formalin-fixed bacteria (Y. ruckeri, S. diarizonae, E. coli O138, O149:F4 and F18+) in separate ELISA plates, and incubated with ppIgG added in serial 2-fold dilutions, and the bound IgG was estimated by using a HRP-conjugated rabbit anti-pig IgG antibody. Data from one typical experiment are presented. (C) ppIgG was tested for reactivity against Escherichia coli and Salmonella diarizonae by competitive ELISA as described ( Materials and Methods ). ppIgG heated at 60°C or 70°C for 1 hour and PBS were used as control samples. The results are presented as ‘% inhibition’ of the signal obtained in the absence of ppIgG (median values (± median ranges)). Two-way ANOVA (subjected to Bonferonni post-test) was used to observe statistical significance between the treated and untreated ppIgG (***: p
Article Snippet: Then, membranes were incubated with polyclonal biotinylated rabbit anti-Pig F(ab)2 IgG (LSBio, Copenhagen, Denmark) diluted 1:10.000 in TBS-T for 1 hour followed by 3x10 min washes in TBS-T before the last incubation with alkaline phosphatase-conjugated streptavidin (DAKO, Glostrup, Denmark) 1:3000 in TBS-T (1 hour).
Techniques: SDS Page, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Competitive ELISA, Inhibition