All lanes : Anti-MBD3 antibody (ab16057) at 0.11 µg/mlLane 1 : HeLaLane 2 : Hela nuclear Lane 3 : A431Lane 4 : HeLa with Human MBD3 peptide (ab16210) at 1 µg/mlLane 5 : HeLa nuclear with Human MBD3 peptide (ab16210) at 1 µgLane 6 : A431 with Human MBD3 peptide (ab16210) at 1 µg/mlSecondaryAlexa Fluor Goat polyclonal anti-Rabbit IgG (700)
ICC/IF image of ab16057 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab16057, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
All lanes : Anti-MBD3 antibody (ab16057) at 1/1000 dilutionLane 1 : Mouse embryonic stem cell lysate at 30 µgLane 2 : Mbd3 null mouse embryonic stem cell lysate at 30 µgLane 3 : MarkerSecondaryGoat anti-rabbit IgG HRP at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.