ICC/IF image of ab46780 stained T24/83 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 ab46780 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- chicken (ab150173) IgY (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
All lanes : Anti-TGF beta 1 antibody (ab46780) at 1/2000 dilution (ab50036, 0.1ug)Lane 1 : As aboveLane 2 : As aboveSecondaryGoat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.