Alexa Fluor® 488 Anti-CD16+CD32 antibody [93]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-CD16+CD32 antibody [93] (AB256417)

ab256417 staining CD16+CD32 in Raw264.7 cells. The cells were fixed with 100% methanol (5 min), 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 ab256417 at 1/500 dilution (shown in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labeled 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 Raw264.7 cells fixed with 4% formaldehyde (10 min).

• Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Anti-CD16+CD32 antibody [93] (AB256417)

Flow cytometry staining of C57 BL/6 mouse splenocytes stained with ab256417 (right) or Rat IgG2aκ Alexa Fluor^® 488 isotype (left). Cells were incubated for 30 min on ice in 1x PBS containing 10 % mouse serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab256417) or Rat IgG2aκ Alexa Fluor^® 488 isotype (1x 10⁶ in 100 μl at 0.2 μg/ml (1/500)) for 30 min on ice. The cells were simultaneously stained with CD19.

Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live single cells.