Cd68 sr d1 antibody

Mouse thyroid tissues were prepared as 4 μm paraffin sections. After antigen retrieval, each section was treated with 100 μL of 3% H₂O₂ for 10 min at room temperature and blocked with 100 μL of 2.5% goat serum blocking solution for 30 min at room temperature. For immunohistochemistry analysis, after removal of the blocking solution, each section was incubated first for 1 h with 100 μL of diluted primary antibodies (IL-17A antibody, 1:500, Cat # NBP1-76337, Novus; CD68/SR-D1 Antibody, 1:150, Cat # NB100-683, Novus) then with 100 μL of secondary antibody for 30 min at room temperature. The positive cells were revealed by incubating with 100 μL of freshly prepared 3,3’-diaminobenzidine (DAB) solution. For immunofluorescence analysis, after removal of the blocking solution, each section was incubated first with 100 μL of diluted primary antibody (IL-17A antibody, 1:400, Cat # NBP1-76337, Novus) overnight at 4°C then with 100 μL of secondary antibody for 2 h at room temperature in the dark. Cells were counterstained with 100 μL 4′,6-diamidino-2-phenylindole (DAPI) for 10 min in the dark.

Pancreas sections from human donors and human IAPP (hIAPP) transgenic mice were stained for macrophages using CD68/SR-D1 antibody (Novus Biologicals, Littleton, CO, USA), or using guinea pig anti-insulin (DAKO) mouse anti-C4BP, sheep anti-pancreatic polypeptide (Bio-Rad) or Congo Red. See ESM Methods for further details.