All lanes : Anti-WNK1 antibody (ab35069) at 1/250 dilutionLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate (ab7900)Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.
ICC/IF image of ab35069 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab35069, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence. 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).