The a5 antibody was raised against the collagenase-resistant residue of human glomerulus basement membrane. It was screened by ELISA and SDS-PAGE to recombinant human type IV collagen chains and further characterized by indirect immunofluorescence within human kidney sections. This detects the bovine a5 chain of type IV collagen (J. Wieslander, personal communication). All of these specific primary antibodies were diluted to 1:100 in antibody diluent (291 mM NaCl, 7.54 mM Na2HPO4, 2.50 mM NaH2PO42H2O, 0.01% NaN3, pH 7.1). The rat monoclonal antibodies were used under denaturing and nondenaturing conditions, whereas the mouse monoclonal antibodies were used only under nondenaturing conditions. birth control yasmin
Rabbit anti-human von Willebrand factor (factor VIII-related antigen) was obtained commercially (cat. #F-3520; Sigma, 1:50 in antibody diluent). Rabbit antiserum to 3p-hydroxysteroid dehydrogenase (3 (3-HSD) type II was kindly donated by J.I. Mason (Clinical Biochemistry, University of Edinburgh; 1:50 in antibody diluent); this antiserum was raised against human placental type I 3p-HSD, but it cross-reacts with type II 3^-HSd, as type I 3p-HSD is 95% homologous to the type II 3p-HSD that is expressed in gonads and the adrenal. Normal sera used in this study for control sections were normal rabbit serum (cat. #R9133; Sigma), normal mouse serum (cat. #M5905; Sigma), normal rat serum (cat. #R9759; Sigma), and normal donkey serum (cat. #D9663; Sigma).