All lanes : Anti-CACNA1C antibody [L57/46] (ab84814) at 1 µg/ml

Lane 1 : Molecular weight marker
Lane 2 : Cell lysates prepared from DHPR alpha 1 transfected CHO cells

ab84814 staining CACNA1C in human hippocampus tissue section by Immunohistochemistry (Bouin's fixative fixed paraffin embedded tissue sections). Tissue underwent heat mediate fixation in microwave and in citrate buffer. The primary antibody was used at 1/100 dilution. A Fluorophore conjugated goat anti-mouse was used as secondary at 1/50 dilution.

ab84814 staining CACNA1C in mouse brain tissue section by immunohistochemistry (Bouin's fixed paraffin embedded tissue section. Tissue underwent heat mediated fixation in microwave and in citrate buffer. The primary antibody was used at 1/100 dilution. A HRP conjugated secondary was used at 10 dilution.

Overlay histogram showing SH-SY5Y cells stained with ab84814 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab84814, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween for 20 min used under the same conditions.

Formaldehyde-fixed SK-N-BE cells stained for CACNA1C (green) using ab84814 at 1/100 dilution in ICC/IF.

Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1/200 dilution for 60 minutes at room temperature. Counterstain: Phalloidin Texas Red F-Actin stain (A); DAPI (B, blue) nuclear stain at 1/1000, 1/5000 for 60 minutes at room temperature, 5 minutes at room temperature.