Immunofluorescent analysis of AKAP9 in A431 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a AKAP9 monoclonal antibody (ab32679) at a dilution of 1:100 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. AKAP9 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunofluorescent analysis of AKAP9 in NIH-3T3 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a AKAP9 monoclonal antibody (ab32679) at a dilution of 1:100 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. AKAP9 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunohistochemistry was performed on normal biopsies of deparaffinized Human pancreas tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a AKAP9 monoclonal antibody (ab32679) at a dilution of 1:20 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.