All lanes : Anti-Alpha B Crystallin (phospho S59) antibody (ab103282) at 1 µg/mlLane 1 : Spinal Cord (Human) Tissue Lysate - adult normal tissue (ab29188)Lane 2 : Spinal Cord (Human) Tissue Lysate - adult normal tissue (ab29188) with Immunising peptide at 1 µg/mlLane 3 : Spinal Cord (Human) Tissue Lysate - adult normal tissue (ab29188) with Non modified peptide at 1 µg/mlLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ab103282 stained HeLa 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 ab103282 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.