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 image of ab25235 stained RAW246.7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab25235, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab98420, DyLight® 488 goat anti-rat IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

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