Alexa Fluor® 594 Anti-CD45 antibody [EP322Y]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 594 Anti-CD45 antibody [EP322Y] (AB303598)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab303598 (right) or Rabbit IgG (monoclonal) Alexa Fluor® 594 (ab208568) isotype (left). Cells were fixed and permeabilized with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 min on ice in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab303598) or Rabbit IgG (monoclonal) Alexa Fluor® 594 (ab208568) isotype (100 μl at 0.2μg/ml (1/2500)) for 30 min on ice.

Acquisition of >30000 events were collected using a 50 mW Yellow/Green laser (561 nm) and 610/20 bandpass filter. Events were gated on live cells.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 594 Anti-CD45 antibody [EP322Y] (AB303598)

Flow cytometry overlay histogram showing left Jurkat positive cells, and right, negative HEK293T cells stained with ab303598 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab303598) (1x 106 in 100μl at 0.04μg/ml (1/12500)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor® 594 (ab208568) 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 Yellow/Green laser (561 nm) and 610/20 bandpass filter.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 594 Anti-CD45 antibody [EP322Y] (AB303598)

ab303598 staining CD45 in Jurkat (Human T cell leukemia cell line from peripheral blood) cells (positive control) and HEK293 cells (negative control). The cells were fixed with 100% methanol (10 min) permeabilized with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3 M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab303598  at 5 µg/ml (shown in red) and ab195883 (Alexa Fluor® 488 Anti-Tubulin antibody [YOL1/34]) at 5 µg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.