Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric (intracellular) analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HEK-293 (human embryonic kidney epithelial cell, left) / Ramos (human Burkitt's lymphoma B lymphocyte, right) cell lines labelling CD20 with ab312151 at 1/5000 dilution (red) compared with Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor^® 555) (ab208569) (black) and an unlabelled control (cell without incubation with primary antibody and secondary antibody) (blue).

10% Goat serum was used as a blocking agent.

Negative control : HEK-293.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MOLT-4(human lymphoblastic leukemia T lymphoblast) cells labelling Notch1 with ab315420 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) (Left panel) / A375 (human malignant melanoma epithelial cell) (Right panel) cells labeling SOX10 with ab307763 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Negative control : HeLa.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometry overlay histogram showing HCT116 cells stained with ab314297 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab314297) (1x 10⁶ in 100μl at 5.0μg/ml (1/100)) for 30min at 22°C.

Isotype control antibody (black line) was Recombinant Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab208569) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Yellow/Green laser (561nm) and 585/42 bandpass filter.

This antibody gave a positive signal in HCT116 fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• Flow Cyt

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Flow Cytometry - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CXCL16 with ab324079 at 1/50 dilution (1ug) / Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) / Left isotype control.

Cells were co-stained with CD19 conjugated Alexa Fluor®647.
Gated on viable cell.

• Flow Cyt

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Flow Cytometry - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CXCL16 with ab324079 at 1/50 dilution (1ug) / Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) / Left isotype control.

Cells were co-stained with CD11b conjugated BV421.
Gated on viable cell.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

• Flow Cyt

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Flow Cytometry - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling TREM1 with ab316942 at 1/50 dilution (1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Human PBMC are co-stained with CD3 conjugated Alexa Fluor®647.
Gated on viable cell.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MDA-MB-231 (human breast adenocarcinoma epithelial cell, Left) / MCF7 (human breast adenocarcinoma epithelial cell, Right) cells labelling CD24 with ab315293 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Negative control : MDA-MB-231.

• Flow Cyt

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Flow Cytometry - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of Isotype (Left) / 293T cells transfected with a human EMR1/ADGRE1 expression vector containing a myc tag (Middle) / 293T cells transfected with an empty vector containing a myc tag (Right) cells labelling EMR1/ADGRE1 with ab316877 at 1/50 dilution (1 ug)/Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cell.

• Flow Cyt

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Flow Cytometry - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling TREM1 with ab316942 at 1/50 dilution (1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Human PBMC are co-stained with CD11b conjugated BV421.
Gated on viable cell.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa(human cervical adenocarcinoma epithelial cell) cells labelling Notch1 with ab315420 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized C6 (rat glial tumor glial cell) cells labeling Bcl-XL with ab306571 at 1/50 dilution (1 ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized B16F0 (mouse melanoma epithelial cell-like) cells labeling SOX10 with ab307763 at 1/50 dilution (1.0µg) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Bcl-XL with ab306571 at 1/50 dilution (1 ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 555) (ab208569) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

• Flow Cyt

Lab

Flow Cytometry - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Overlay histogram showing HeLa cells stained with ab202520 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab202520, 1/50 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 555 (ab208569) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB208569)

Immunofluorescent analysis of HeLa (human cervical cancer) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 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 ab208569 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/1000 dilution (showing no signal) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).