Journal: Journal of Alzheimer's Disease

Article Title: One of the binding proteins for administered amyloid-β appears to be anti-Aβ IgG antibody in amyloid plaques

doi: 10.1177/13872877251389631

Figure Lengend Snippet: Specific binding of applied Bio42 to APs demonstrated by a tissue competition assay (A-I) and schematic summary of the results (J, K). (A) Labeling of APs (arrowheads) with Bio42 in a brain tissue from a dementia and CAA patient is presented. Binding of Bio42 to APs and artery walls (asterisks) was specific to Aβ 42 , as coincubation with non-biotinylated Aβ 42 peptide (Pep42) progressively attenuated the binding in dose-dependent manner (Pep42/Bio42 ratio: 0.01 for (B); 0.1 for (C); 1 for (D); and 10 for (E)). (F) No labeling was observed without Bio42 as a negative control . In contrast, coincubation with non-biotinylated Aβ 40 peptide (Pep40) had little effect on Bio42 binding (Pep40/Bio42 ratio: 0.1 for (G); 1 for (H); and 10 for (I)), unlike Pep42 (scale bar, 100 μm). A fixed amount of Bio42 (1 μg/ml) was coincubated stepwise with Pep42 (J) or Pep40 (K) in various ratios. The corresponding AP labeling results shown in (A-E) with Pep42 and in (G-I) with Pep40 are summarized in tables. AP labeling with Bio42 was dense in the absence of Pep42 but gradually became fainter as the Pep42/Bio42 ratio increased. Given that Bio42 and Pep42 specifically bind to the BP and that Pep40 is noncompetitive, dense AP labeling is expected to be invariably preserved, regardless of the Pep40/Bio42 ratio.

Article Snippet: The following well-characterized antibodies were used at the appropriate concentrations for immunohistochemical, immunofluorescence staining, or WB analyses: biotinylated anti-human IgG antibody (Vector Laboratories), anti-biotin antibody (Jackson ImmunoResearch), goat anti-human IgG antibody (Merck).

Techniques: Binding Assay, Competitive Binding Assay, Labeling, Negative Control

Journal: Journal of Alzheimer's Disease

Article Title: One of the binding proteins for administered amyloid-β appears to be anti-Aβ IgG antibody in amyloid plaques

doi: 10.1177/13872877251389631

Figure Lengend Snippet: Labeling of amyloid plaques (APs) by biotinylated synthetic Aβ 42 peptide (Bio42) visualized by diaminobenzidine (DAB). (A-D) Brain sections including APs and cerebral amyloid angiopathy (CAA) were incubated with 0.1 μg/ml, 1 μg/ml, 10 μg/ml, and 100 μg/ml of Bio42 and stained by DAB. Three representative arteries are marked as landmarks (asterisks) that appear in all serial sections. (A) APs and artery walls were labeled clearly with 0.1 μg/ml of applied Bio42. (B) With 1 μg/ml of applied Bio42 the same APs and artery walls were labeled more densely with higher background. In addition, a larger number of APs were observed (arrowheads). (C) In contrast, with 10 μg/ml of applied Bio42 the labelling density was markedly decreased. (D) Moreover, the labeling was absent with 100 μg/ml of applied Bio42. There is an optimal concentration of binding for applied Bio42 to APs (scale bar, 100 μm).

Article Snippet: The following well-characterized antibodies were used at the appropriate concentrations for immunohistochemical, immunofluorescence staining, or WB analyses: biotinylated anti-human IgG antibody (Vector Laboratories), anti-biotin antibody (Jackson ImmunoResearch), goat anti-human IgG antibody (Merck).

Techniques: Labeling, Incubation, Staining, Concentration Assay, Binding Assay