All lanes : Anti-Slit1 antibody (ab88491) at 1/200 dilutionLane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLane 2 : HEK293 cells expressing mSlit1-mycLane 3 : HEK293 cells expressing hSlit2-mycLane 4 : HEK293 cells expressing hSlit3-mycLysates/proteins at 30 µg per lane.SecondaryHRP Anti-Rabbit at 1/20000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab88491 stained HepG2 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 (ab88491, 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.