All lanes : Anti-IGF1 Receptor (phospho Y1161) antibody (ab85625) at 1 µg/mlLane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate with Human IGF1 Receptor (phospho Y1161) peptide (ab99365) at 1 µg/mlLane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Human IGF1 Receptor (phospho Y1161) peptide (ab99365) at 1 µg/mlLane 3 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330) with Human IGF1 Receptor (phospho Y1161) peptide (ab99365) at 1 µg/mlLane 4 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate with Human IGF1 Receptor peptide (ab118178) at 1 µg/mlLane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Human IGF1 Receptor peptide (ab118178) at 1 µg/mlLane 6 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330) with Human IGF1 Receptor peptide (ab118178) at 1 µg/mlLane 7 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate Lane 8 : HEK293
ICC/IF image of ab85625 stained MCF-7 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 ab85625 at 5µg/ml overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.