Anterior lens capsules with attached lens epithelial cells were
fixed in 10% neutral-buffered formalin and embedded in paraffin. The
paraffin-embedded samples were sectioned on a microtome at a
thickness of 5 μm, deparaffinized in xylene, and rehydrated in
alcohol. The sections were incubated in 2%
H2O2 for 5 minutes, 20%
nonimmune horse serum (Biomeda, Foster City, CA) for 10 minutes, and
1:200 dilution of rabbit anti–βig-h3 antiserum or normal rabbit
serum for 2 hours at room temperature. The sections were then incubated
in biotinylated anti-rabbit IgG (Amersham, Cleveland, OH) for
10 minutes, and then visualized according to the manufacturer’s
protocol using a detection kit (UltraTek HRP; ScyTek Laboratories,
Logan, UT). The immunolabeled sections were counter-stained with 10%
Mayer’s hematoxylin and examined under light microscope.