All lanes : Anti-TRIM21 antibody (ab91423) at 0.04 µg/mlLane 1 : HeLa whole cell lysate at 50 µgLane 2 : HeLa whole cell lysate at 15 µgLane 3 : HeLa whole cell lysate at 5 µgdeveloped using the ECL technique
SSA1 was immunoprecipitated from 1mg HeLa whole cell lysate using 10µg ab91423. 20% of the immunoprecipitate was loaded per lane, and probed with ab91423 at 1µg/ml (lane 1) or a control IgG (lane2). Detection: chemoluminescence with an exposure time of 10 seconds.
ICC/IF image of ab91423 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab91423, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit 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.