All lanes : Anti-GRO alpha antibody (ab86436) at 0.5 µg/mlLane 1 : Jurkat cell lysateLane 2 : 3T3 cell lysate
ICC/IF image of ab86436 stained HeLa cells. The cells were 4% formaldehyde (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 (ab86436, 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.
All lanes : Anti-GRO alpha antibody (ab86436) at 0.5 µg/mlLane 1 : Active GRO alpha full length protein (ab9773) at 0.1 µgLane 2 : Active GRO alpha full length protein (ab9773) at 0.01 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 1 minute