Anti-CaV1.3 antibody [L48A/9] (ab85491)

ab85491 staining CaV1.3 in human hippocampus tissue by IHC-P (Bouin's fixed paraffin embedded tissue sections).

The primary antibody was used at 1:1000 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.

ab85491 staining CaV1.3 in human differentiated iPS cells by Immunocytochemistry/ Immunofluorescence.

The cells were fixed in paraformaldehyde, permeabilized in 0.05% Tween-20 and then blocked using 3% BSA for 1 hour at 21°C. Samples were then incubated with primary antibody at a 1/100 dilution for 12 hours at 4°C. The secondary antibody used was a anti-mouse IgG (H+L) conjugated to Alexa Fluor^® 594 (pink) used at a 1/400 dilution.

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Overlay histogram showing SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells stained with ab85491 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab85491, 0.5 µg/1x10⁶ cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1 µg/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 minutes)/permeabilized in 0.1% PBS-Tween for 20 minutes used under the same conditions.

ab85491 staining CaV1.3 in mouse backskin tissue by IHC-P (Bouin's fixed paraffin embedded tissue sections).

Tissue underwent antigen retrieval using microwave in citrate buffer. The primary antibody was used at 1/100 dilution and then sections were incubated with Fluorophore conjugated goat anti mouse at 1/50 dilution.