Overlay histogram showing HepG2 cells stained with ab18180 (red line). The cells were fixed with 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 (ab18180, 2µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

Western blot analysis of ABCA1 induction by cholesterol (+) in human fibroblast cells from different patients (1,2,3,4). Simultaneous bloting with anti-actin antibody was used for protein loading control. Western blot analysis of ABCA1 induction by cholesterol (+) in human fibroblast cells from different patients (1,2,3,4). Simultaneous blotting with anti-actin antibody was used for protein loading control.

All lanes : Anti-ABCA1 antibody [AB.H10] (ab18180) at 1/1000 dilution

Lanes 1 & 3 & 5 : Wild type mouse liver tissue lysate
Lanes 2 & 4 & 6 : Knockout mouse liver tissue lysate

Lysates/proteins at 100 µg per lane.

Predicted band size: 254 kDa
Observed band size: 254 kDa

The lower bands are from the secondary anti-mouse antibody reacting to the endogenous mouse antibodies from the tissue.

Immunoreactivity of pancreas tissue for ABCA1 protein. Left panel: strong immunopositivity of exocrine glandular cells of the pancreas, especially in the basal region (arrows). Interstitial microvascular cells are apparently negative. Cells within the islet were weakly positive (data not shown). Right panel: the negative control. Hematoxylin counterstain.