The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Detects a band of approximately 101 kDa (predicted molecular weight: 101 kDa).
We recommend using 3% milk as the blocking agent in Western Blot.
Use 1-2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ORP8 is a member of the oxysterol-binding protein related protein family. In humans, the gene family consists of 12 members, and extensive splice variation increases the number of encoded protein products substantially. The ORPs have been implicated as sterol sensors that regulate a number of cellular functions including sterol and neutral lipid metabolism, intracellular lipid transport, membrane trafficking and cell signaling. ORP8 has been recently shown to act as a sterol sensor that affects the reverse cholesterol transport process via modulation of ABCA1 expression and macrophage cholesterol efflux.
Overlay histogram showing HEK293 cells stained with ab60110 (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 (ab60110, 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 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 HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.