Recombinant Anti-CD105 antibody [MJ7/18] (ab81456)

Lanes 1 - 3: Merged signal (red and green). Green - ab81456 observed at 80 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

ab81456 was shown to react with CD105 in Western blot.

Membranes were blocked in 3% milk before incubation with ab81456 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 5 µg/ml and a 1 in 20000 dilution respectively.

Blots were incubated with Goat anti-Rat IgG H&L (IRDye® 800CW) preabsorbed (ab253031) and Goat anti-Rabbit IgG H&L (IRDye$®$ 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

Additional band(s) were observed at 45 kDa, these band(s) may represent alternative splice variants. This has not been investigated further.

Lane 1: Mouse Lung tissue lysate (40 µg)
Lane 2: Mouse Liver tissue lysate (40 µg)
Lane 3: NIH/3T3 whole cell lysate (40 µg)

Flow cytometry overlay histogram showing left bEND.3 positive cells and right negative NIH3T3 cells stained with ab81456 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab81456) (1x10⁶ in 100 μl at 1 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 for 30 min on ice.

Isotype control antibody (black line) was Rat IgG2aκ (ab18450) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.