Anti-CD11c antibody [3.9]

• Flow Cyt

Lab

Flow Cytometry - Anti-CD11c antibody [3.9] (AB11029)

Human peripheral blood lymphocytes stained with ab11029. Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID : 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lysed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were stained with anti-CD11c ab11029 (right panel) at 1/100 dilution for 30 min at 4°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-mouse IgG (H&L) (ab150117) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (left panel) was mouse monoclonal IgG1 (ab170190) used under the same conditions.

Acquisition of >30,000 total events were collected using a 50mW Argon Blue laser (488nm) and 530/30 bandpass filter. Gating strategy – events were collected with the forward and side light-scatter characteristics of viable cells.

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD11c antibody [3.9] (AB11029)

Human whole blood stained with ab11029 (right) or mouse IgG1κ (left). Red blood cells of 200μl human whole blood were lysed, then cells 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 (ab11029) or mouse IgG1κ isotype (ab170190) (100μl at 10 μg/ml) for 30 min on ice.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ^® 488, pre-adsorbed) (ab150177) was used at 1/2000 dilution for 30 min at 4°C.

Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on viable single cells.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD11c antibody [3.9] (AB11029)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab264107 (right) or Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab264107 or Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (1x 10⁶ in 100 µl at 0.2 μg/ml (1/5000)) for 30min on ice. The cells were simultaneously stained with CD14.

The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS only (ab264107).