Journal:

Article Title: Isolation and Characterization of Two Proteins from Moraxella catarrhalis That Bear a Common Epitope

doi:

Figure Lengend Snippet: Interaction of proteins with fibronectin and vitronectin determined by dot blot. (A) Binding of vitronectin to UspA1 and UspA2. For this blot, the purified proteins were spotted onto the membrane, blocked, and incubated with human plasma vitronectin. The bound vitronectin was detected with a rabbit antivitronectin conjugate. (B) Binding of fibronectin to UspA1. Purified fibronectin was spotted onto the membrane, blocked, and incubated with purified UspA1. The bound UspA1 was then detected with MAb 17C7. (C) Binding of fibronectin to UspA2. This procedure was done as described for panel B except with UspA2. The amount of protein applied at each spot is indicated on the left.

Article Snippet: After the proteins had been absorbed into the membrane, it was blocked with BLOTTO for 1 h and incubated with human plasma vitronectin (1 μg/ml in BLOTTO; GIBCO BRL, Grand Island, N.Y.) for 4 h at room temperature.

Techniques: Dot Blot, Binding Assay, Purification, Incubation

Journal:

Article Title: Chimeric Plant Virus Particles Administered Nasally or Orally Induce Systemic and Mucosal Immune Responses in Mice

doi:

Figure Lengend Snippet: Intranasal immunization with CPMV-MAST1 without adjuvant elicits FnBP-specific antibody that inhibits binding of fibronectin to GSTD1-3. Mice were immunized intranasally (i.n.) with 100 μg of CPMV-MAST1 on days 0, 7, 14, and 21. Serum was collected on day 50, and the IgG fraction was tested for the ability of the GSTD1-3-specific antibody to inhibit the binding of 5 mg of human plasma fibronectin per ml to immobilized GSTD1-3. Sera from mice taken 42 days after subcutaneous (s.c.) immunization with 100 μg of CPMV-MAST1 in QS-21 on days 0, 14, and 28 were tested for comparison. The IgG from mice immunized with CPMV expressing an unrelated peptide (CVP control) and normal mouse IgG were tested as negative controls. Results are expressed as the percent inhibition of binding achieved with the different IgG samples compared to that in the absence of IgG (0%).

Article Snippet: Human plasma fibronectin (Sigma) was then added to the wells (50 μl/well) at a final concentration of 5 μg/ml and incubated for 1 h at 37°C.

Techniques: Binding Assay, Expressing, Inhibition