ab5595 staining alpha A Crystallin in non-transgenic 13.5 mouse embryos by Immunohistochemistry (PFA fixed, paraffin embedded sections).Heads of mice were removed, fixed in 10% formalin, dehydrated, embedded in paraffin, sectioned (5-7 µm).Slides containing ocular sections from non-transgenic embryos were first deparaffinized and rehydrated. Antigens were retrieved by microwave treatment in 10 mM Sodium Citrate buffer (pH 6.0). Following antigen retrieval, the tissue sections were blocked with 10% normal horse serum for 30 minutes, at room temperature. The slides were then incubated with ab5595 at a 1/10,000 dilution overnight at 4°C. Following brief washes in PBS, the slides were incubated with the appropriate biotinylated-secondary antibodies for 30 minutes at 37°C. Antigen-antibody complexes were then detected using streptavidin-linked Alexa 594 (red) at 1/1000 dilution. Sections were mounted using anti-fade media containing DAPI (blue). Images were captured using a Nikon Eclipse E6