All lanes : Anti-Bad antibody (ab90435) at 1/1000 dilutionLane 1 : Cow normal mammary epithelial cells (nMEC) treated with serum free mediaLane 2 : Cow normal mammary epithelial cells (nMEC) treated with 1.0µM anisomycinLane 3 : Cow normal mammary epithelial cells (nMEC) treated with 100ng/ml IGF-1Lane 4 : Cow normal mammary epithelial cells (nMEC) treated with anisomycin and IGF-1Lane 5 : Cow normal mammary epithelial cells (nMEC) treated with serum free mediaLane 6 : Cow normal mammary epithelial cells (nMEC) treated with 1.0µM anisomycinLane 7 : Cow normal mammary epithelial cells (nMEC) treated with 100ng/ml IGF-1Lane 8 : Cow normal mammary epithelial cells (nMEC) treated with anisomycin and IGF-1Lysates/proteins at 40 µg per lane.SecondaryECL Anti-Rabbit IgG, Horseradish Peroxidase Linked at 2500 cellsdeveloped using the ECL techniquePerformed under reducing conditions.
Anti-Bad antibody (ab90435) at 1 µg/ml + Mouse Raw 264 cell lysatedeveloped using the ECL technique
ab90435, at a 1/50 dilution, staining BAD in the neurons of paraffin embedded Alzheimer diseased brain tissue (right) by Immunohistochemistry. Control brain tissue (left).
ICC/IF image of ab90435 stained RAW246.7 cells. The cells were 4% paraformaldehyde 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 (ab90435, 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.