ICC/IF image of ab136550 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 ab136550 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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.
All lanes : Anti-CD47 antibody (ab136550) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell LysateLane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 4 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell LysateLane 5 : THP1 (Human acute monocytic leukemia cell line) Whole Cell LysateLane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunising peptide at 1 µg/mlLane 7 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunising peptide at 1 µg/mlLane 8 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Immunising peptide at 1 µg/mlLane 9 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate with Immunising peptide at 1 µg/mlLane 10 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate with Immunising peptide at 1 µg/mlLysates/proteins at 25 µg per lane.S