重组Anti-HLA Class I抗体[W6/32] (ab22432)

Immunohistochemical analysis of ab22432 10% paraformaldehyde fixed endothelial cells in frozen Human spleen tissue Human heart tissue labeling HLA Class I with ab22432 at 0.05µg/ml. Detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% PBS-Tween permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling HLA Class I with ab22432 at 1µg/mL, 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 ab92494 at 1µg/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 red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% PBS-Tween permeabilized negative cell line K562 labelling HLA Class I with ab22432 at 1µg/mL, 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 ab227805 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 red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

Flow cytometry overlay histogram showing left Jurkat positive cells and right negative K562 cells stained with ab22432 (red line). The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab22432) (1x10⁶ in 100 μl at 0.2 μg/ml) for 30 min on ice. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at for 30 min on ice. Isotype control antibody (black line) was mouse IgG2aκ (ab18413) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.