Brefeldin A, Inhibitor of ADP-ribosylation factor

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Brefeldin A, Inhibitor of ADP-ribosylation factor (AB120299)

ab84340 staining golgin-97 in MCF7 cells treated with brefeldin A (ab120299), by ICC/IF. Increase in Golgin-97 expression correlates with increased concentration of brefeldin A, as described in literature.
The cells were incubated at 37°C for 1.5 h in media containing different concentrations of ab120299 (brefeldin A) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab84340 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Brefeldin A, Inhibitor of ADP-ribosylation factor (AB120299)

ab66579 staining TNF alpha in Raw264.7 cells. The cells were treated with LPS for 7 hours and Brefeldin A (ab120299) for the final 3 hours. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab66579 at 1μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• Chemical Structure

Lab

Chemical Structure - Brefeldin A, Inhibitor of ADP-ribosylation factor (AB120299)

2D chemical structure image of ab120299, Brefeldin A, Inhibitor of ADP-ribosylation factor