Journal: Experimental Animals

Article Title: Experimental transmission of systemic AA amyloidosis in autoimmune disease and type 2 diabetes mellitus model mice

doi: 10.1538/expanim.16-0037

Figure Lengend Snippet: Amyloid deposits in spleens of several mice. Amyloid deposits were stained with Congo red and observed as green birefringence under polarized light. Amyloid fibrils in spleens of 18-week-old mice (C57BL, C57BL lpr/lpr , C57BL Ham +/+, and C57BL ob/ob ) inoculated with murine amyloid fibrils were reacted with AF2948 antibody in IHC. Also, amyloid fibrils in spleens of 18-and 30-week-old C57BL Ham +/+ mice inoculated with bovine amyloid fibrils and chicken amyloid fibrils, respectively, were reacted with AF2948 antibody in IHC. Bar=100 µ m.

Article Snippet: The membranes were incubated with anti-mouse SAA1 goat antibody (1:5,000) (AF2948, R&D Systems, Minneapolis, MN, USA), anti-bovine SAA1 monoclonal antibody, 25BF12 (1:1,600) [ ], which was purified from the culture supernatant of hybridoma cells by protein A-sepharose (GE Healthcare), or anti-chicken SAA1 mouse antibody (1:1,600) [ ] in 1% skim milk in PBST at room temperature for 1 h. Membranes were washed three times with PBST and incubated with anti-goat IgG donkey antibody F (ab’) 2 conjugated with horseradish peroxidase (HRP) (1:1,000) (SC3851, Santa Cruz Biotechnology, Dallas, TX, USA) , anti-mouse IgG F (ab’) 2 conjugated with HRP (1:1,600) (NA9310V, GE Healthcare, Buckinghamshire, UK) in 1% skim milk in PBST at room temperature for 45 min.

Techniques: Staining

Journal: Experimental Animals

Article Title: Experimental transmission of systemic AA amyloidosis in autoimmune disease and type 2 diabetes mellitus model mice

doi: 10.1538/expanim.16-0037

Figure Lengend Snippet: Intensity scoring of amyloid deposits in the spleen using ImageJ. Amyloid fibrils in spleens of mice inoculated with murine, bovine, or chicken amyloid fibrils were reacted with AF2948 antibody in IHC. Images of IHC indicate amyloid deposits around spleen follicles. ImageJ was used to measure the intensity of amyloid deposits. ImageJ intensity scores are shown at the bottom. Bar=400 µ m.

Article Snippet: The membranes were incubated with anti-mouse SAA1 goat antibody (1:5,000) (AF2948, R&D Systems, Minneapolis, MN, USA), anti-bovine SAA1 monoclonal antibody, 25BF12 (1:1,600) [ ], which was purified from the culture supernatant of hybridoma cells by protein A-sepharose (GE Healthcare), or anti-chicken SAA1 mouse antibody (1:1,600) [ ] in 1% skim milk in PBST at room temperature for 1 h. Membranes were washed three times with PBST and incubated with anti-goat IgG donkey antibody F (ab’) 2 conjugated with horseradish peroxidase (HRP) (1:1,000) (SC3851, Santa Cruz Biotechnology, Dallas, TX, USA) , anti-mouse IgG F (ab’) 2 conjugated with HRP (1:1,600) (NA9310V, GE Healthcare, Buckinghamshire, UK) in 1% skim milk in PBST at room temperature for 45 min.

Techniques: