Retinae were collected from adult White Leghorn chicken and fixed in the eyecup for two hours in 4% Paraformaldehyde (in PBS). Retinae were then removed from the eyecups and incubated in 30% sucrose (in PBS) overnight. Retinal tissue was embedded and frozen in OCT compound and cut into ~15um sections on a cryotome. Sections were blocked in 5% normal goat serum (in 1%BSA/.1%saponin PBS) for one hour and then incubated at RT with 1:250 (1%BSA/.1% saponin PBS) ClC4 antibody for 1 hour. Sections were then washed in PBS (3X10minutes) and then treated with secondary antibody (1:500 Cy3) for one hour. After another PBS wash series, sections were coverslipped and antibody labeling was visualized at 20X with a Leica upright microscope using a TRITC filter set and Xenon LAMP illumination. (Crousillac et al, 2003)
Immunofluorescence image of cultured chick retinal amacrine (neuronal) cells labeled with CLC4 Antibody. Data courtesy of Emily McMains, Louisiana State University.
Western blot of chicken brain tissue incubated with CLC4 Antibody. Data courtesy of Emily McMains, Louisiana State University.