Anti-CD32B + CD32A antibody [AT10]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD32B + CD32A antibody [AT10] (AB23336)

Overlay histogram showing K562 cells stained with ab23336 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23336, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 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 result in 4% paraformaldehyde (10 min) fixed K562 cells used under the same conditions.

Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD32B + CD32A antibody [AT10] (AB23336)

ab23336 staining CD32B + CD32A in K-562 cells. The cells were fixed with 4% paraformaldehyde (10 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 ab23336 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

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