Anti-CD16+CD32 antibody [93] - BSA and Azide free

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD16+CD32 antibody [93] - BSA and Azide free (AB25235)

C57BL/6 mouse splenocytes stained with ab25235 (right) or Rat IgG2aκ (ab18450) isotype (left). C57BL/6 mouse splenocytes were incubated for 30 min on ice in PBS / 10 % mouse serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab25235) or Rat IgG2aκ (ab18450) (1x10⁶ in 100μl at 5 μg/ml) for 30 min on ice.

The secondary antibody Goat anti-rat IgG H&L (Alexa Fluor ^® 488, pre-adsorbed) (ab150165) was used at 1/2000 dilution for 30 min at 4°C. The cells were simultaneously stained with CD19 antibody.

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

This image was generated using the ascites version of the product.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD16+CD32 antibody [93] - BSA and Azide free (AB25235)

ab25235 staining CD16 + CD32 in Raw264.7 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween 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 ab25235 at 10µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150165, Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.