Anti-CACNA1S antibody [1A] (ab2862)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) was performed on normal biopsies of deparaffinized human skeletal muscle tissue. Heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then incubated with ab2862 (1:20) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively 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.

Flow cytometry analysis of Dihydropyridine Receptor alpha-1 in U251 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5 x10⁶ cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Dihydropyridine Receptor alpha-1 monoclonal antibody at 1 µg/test for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.