All lanes : Anti-WASP antibody (ab74904) at 1 µg/mlLane 1 : Saos 2 (Human epithelial-like osteosarcoma cell line) Whole Cell LysateLane 2 : LOVO (Human colon adenocarcinoma cell line) Whole Cell LysateLane 3 : HT 1080 (Human fibrosarcoma) Whole Cell LysateLane 4 : EBS E14 TG2A Day 0 (Mouse Pluripotent Embryonic Stem Cell) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
IHC image of WASP staining in human Cerebral cortex FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab74904, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
All lanes : Anti-WASP antibody (ab74904) at 1 µg/mlLane 1 : Saos-2Lane 2 : LOVO (Human colon adenocarcinoma cell line) Whole Cell Lysate Lane 3 : HT 1080 (Human fibrosarcoma) Whole Cell LysateLane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
ICC/IF image of ab74904 stained HepG2 cells. The cells were 4% PFA 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 (ab74904, 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.