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

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: Analysis of immunohistochemical staining of RAGE and HMGB1 for TETs.

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques: Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: Expression of HMGB1 in thymoma type A (A), B-component type AB (B), B1 (C) is shown. Scale bar: 40 μm. On this example of B1 thymoma lymphocytes are intermingled with few tumor cells. Focus on cytoplasmic staining: a larger magnification of a WHO type B1 thymoma is shown to better display “autophagic” tumor cells (tumor cells with brownish granular cytoplasmic and absent [only hematoxylin blue] nuclear staining). Scale bar: 20 μm (D). Analogously, a type B2 (E) and B3 (F) thymoma are shown. Two examples of TC (SCC (G) and (H)) are displayed – scale bar: 40 μm. HMGB1 high mobility group box1, TETs thymic epithelial tumors, TC thymic carcinoma, SCC squamous cell carcinoma

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques: Expressing, Staining

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: Immunohistochemistry revealed strong expression of RAGE in subcapsular cTEC of fetal (A) and adult thymus (B). Scale bar: 40 μm. For comparison the staining pattern of cytokeratins 5 and 14 – markers of epithelial cell origin on fetal thymus is shown (C). The expression pattern for HMGB1 in cTEC of fetal (D, scale bar: 40 μm) and adult thymus (E, scale bar: 20 μm) is shown. (F) A larger magnification of E is shown. Scale bar 8 μm. Arrows in F indicate HMGB1 cytoplasmic staining in cTEC. RAGE receptor for advanced glycation endproducts, HMGB1 high mobility group box1, cTEC cortical thymic epithelial cells

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques: Immunohistochemistry, Expressing, Comparison, Staining

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: Hassall's corpuscles stained with antibodies to RAGE (A), and HMGB1 (B, small arrows point to small GC) are displayed. Scale bar: 80 μm. Similarly, RAGE (C), and HMGB1 (D) staining in GC of MG patients (scale bar: 80 μm); and RAGE expression in thymic medulla (E, arrows point to thymic medulla; scale bar: 200 μm) and macrophages (F, scale bar: 40 μm) of regular adult thymus are shown. RAGE receptor for advanced glycation endproducts, HMGB1 high mobility group box1, MG Myasthenia gravis, GC germinal center.

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques: Staining, Expressing

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: Concentration of sRAGE, esRAGE and HMGB1 in serum of patients.

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: Levels of sRAGE (a), esRAGE (b) and HMGB1 (c) in sera of patients with TETs including patients with paraneoplastic MG (MG n = 11) compared to healthy volunteers are shown. To rule out the influence of MG on levels of circulating sRAGE (d) and HMGB1 (e) in patients with TETs, patients with MG were excluded from this analysis. The levels of sRAGE in non-invasive (Masaoka-Koga stage I) and invasive TETs (Masaoka-Koga stages II-IV) are shown (f). RAGE receptor for advanced glycation endproducts, sRAGE soluble RAGE, esRAGE endogenous secretory RAGE, HMGB1 high mobility group box1, TETs thymic epithelial tumors, n number, MG Myasthenia gravis.

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques:

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: The levels of circulating sRAGE (a), and HMGB1 (b) in serum of patients with TETs compared to patients with thymic hyperplasia and healthy volunteers are shown. RAGE receptor for advanced glycation endproducts, sRAGE soluble RAGE, HMGB1 high mobility group box1, TET thymic epithelial tumor.

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques:

Journal: PLoS ONE

Article Title: Expression of RAGE and HMGB1 in Thymic Epithelial Tumors, Thymic Hyperplasia and Regular Thymic Morphology

doi: 10.1371/journal.pone.0094118

Figure Lengend Snippet: Patients with TETs were separated into patients with thymomas and TC, and compared to healthy volunteers. Serum concentrations of sRAGE (a) and HMGB1 (b) are shown. Patients with TC were analyzed compared to volunteers for sRAGE (c), esRAGE (d), as well as HMGB1 (e). TETs thymic epithelial tumors, RAGE receptor for advanced glycation endproducts, sRAGE soluble RAGE, esRAGE endogenous secretory RAGE, HMGB1 high mobility group box1, TC thymic carcinoma

Article Snippet: Sections were stained using affinity-purified polyclonal goat anti-human RAGE IgG (R&D Systems, Minneapolis, MN, USA) or monoclonal mouse anti-human HMGB1 IgG 2b (R&D Systems) and biotinylated anti-goat IgG or anti-mouse IgG secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

Techniques: