All lanes : Anti-Glucocorticoid Receptor antibody (ab55400) at 1/500 dilutionLane 1 : Jurkat cell extract, treated with EGF (200ng/ml, 15mins), without immunizing peptideLane 2 : Jurkat cell extract, treated with EGF (200ng/ml, 15mins), with immunizing peptide
ab55400 immunoprecipitating Glucocorticoid Receptor from Hepa1-6 (murine hepatoma) cells. 1 mg of cell lysate was incubated with the primary antibody and matrix (Protein A) at a concentration of 20 µl/mg lysate for 18 hours at 4°C in cell lysis buffer. Immunoprecipitate was then Western blotted with ab55400 at 1/500.See Abreview
ICC/IF image of ab55400 stained PC12 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 (ab55400, 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.