Anti-CD66b antibody [EPR27440-29]

• Flow Cyt

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Flow Cytometry - Anti-CD66b antibody [EPR27440-29] (AB322980)

Flow cytometric analysis of Human purified neutrophils labelling CD66b with ab322980 at 1/5000 dilution (0.01 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD66b antibody [EPR27440-29] (AB322980)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HL-60 (human acute promyelocytic leukemia promyeloblast) cells labelling CD66b with ab322980 at 1/50 (10.24 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing increased cytoplasmic staining in differentiated HL-60 cells (treatment with DMSO (1.25%) for 5 days, shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• Flow Cyt

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Flow Cytometry - Anti-CD66b antibody [EPR27440-29] (AB322980)

Flow cytometric analysis of HL-60 (human acute promyelocytic leukemia promyeloblast) undifferentiated cells (Upper left and right) / HL-60 cells differentiated with 1.25% DMSO for 5 days (Lower left and right) cells labelling CD66b with ab322980 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CD66b antibody [EPR27440-29] (AB322980)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Isotype (Upper Left) / 293T (human embryonic kidney epithelial cell) transfected with an CEACAM8 expression vector containing a myc-His-tag® (Upper Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Upper Right) / 293T cells transfected with homologous human CEACAM expression vector (CEACAM1, CEACAM3, CEACAM5, CEACAM6 ) containing a myc-His-tag® (Lower) cells labelling CD66b with ab322980 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.