重组Anti-CD33抗体[WM53] (ab30371)

ab30371 staining CD33 in HL-60 cells. The cells were fixed with 4% paraformaldehyde (10min), 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 ab30371 at 10μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) 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 pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Flow cytometry staining of human whole blood with ab30371 (right) or mouse IgG1 and kappa; (ab170190) isotype (left). Red blood cells of 200 µL were lysed, then cells were incubated for 30 mins on ice in 1 x PBS containing 10 µg/mL human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interactions followed by antibody (ab30371) or mouse IgG1 & kappa; (ab170190) isotype for 30 min on ice. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor 488, Pre-absorbed) (ab150119) was used at 1/2000 dilution for 30 min on ice. The cells were simultaneously stained with CD14.