Primary mouse anti desmin antibody

Kidney tissues were fixed with buffered 4% paraformaldehyde, embedded in paraffin and cut into 4-μm-thick sections. Periodic acid-Schiff and Masson's trichrome staining were performed using serial sections. Ferric iron deposits were stained using berlin blue staining. Glomerular and tubular lesions were evaluated by a semiquantitative score using the method as previously described. 20 The extent of iron deposits was evaluated by semiquantitative score; 0, 1 and 2 correspond to 0, 1-10 and 10-20% of the microscopic field. Thirty randamly selected fields were counted. Renal sections were immunohistochemically stained with a primary mouse anti-CD68 antibody (AbD Serotec, Raleigh, NC, USA; dilution 1:1000), a primary mouse anti-desmin antibody (Dako, Glostrup, Denmark; dilution 1:50). Immunostains were visualized with the use of EnVision+ System-HRP (DAB) (Dako). Every section was counterstained with hematoxylin. Quantification of CD68-positive cells and desmin staining was evaluated as previously described. 15 All of the histological scoring was performed in a blinded manner.