Hep 2 ana kit

Individual serum from each treatment group was evaluated for the presence of anti-nuclear antibodies using HEp-2 ANA kit (Inova Diagnostics, Inc., San Diego, CA, USA) as previously described [28 (link)–31 (link)]. Manufacturer’s specific directions were followed in all procedures. Each serum sample was diluted 1:80 and incubated on HEp-2-fixed substrate slides for 1 h at room temperature in a humidified chamber. After three 5-min washes with PBS, the substrate slides were treated with a 1:100 dilution of Alexa 488 goat anti-mouse IgG (H + L) (Life Technologies) for 45 min at room temperature. After three washes, Vectashield DAPI mounting medium (Vector Laboratories, Burlingame, CA, USA) was applied, and overlaid with a glass coverslip. Fluorescence was detected by fluorescence microscopy at ×400 magnification by using a Nikon microscope, and all images were obtained with exposure of 200 ms.

Anti-nuclear antibodies were detected in the sera of mice by using HEp-2 ANA kit (Inova Diagnostics, Inc., San Diego, CA, USA). All procedures were performed according to the manufacturer's instructions. Mouse sera were diluted 1:80 and incubated on HEp-2-fixed substrate slides for one hour at room temperature in a humidified chamber. After three 5-min washes with PBS, the substrate slides were treated with a 1:100 dilution of AF 488 goat anti-mouse IgG (H+L) (Life Technologies) for 45 min at room temperature. After three washes, slides were treated with Vectashield DAPI-mounting medium (Vector Laboratories) and overlaid with glass coverslips. Fluorescence was detected by fluorescence microscopy at 400× magnification by using a Nikon microscope, and all images were obtained with exposure of 200 ms.