ICC/IF image of ab21738 stained MCF7 cells. The cells were 100% Methanol fixed (5 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 (ab21738, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 Goat anti-Rabit 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.
IHC image of PLK1 (phospho S137) staining in human normal colon 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 ab21738, 1µg/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
This antibody gives a positive result in ELISA against the modified peptide ab25712 (orange dotted line) but does not recognise the corresponding non modified peptide ab25713 (yellow dotted line). This indicates that ab21738 specifically recognises PLK1 phosphorylated at S137.
PLK1 (phospho S137) immunoprecipitation from Human HeLa cells using ab21738. 200 µg of cell lysate was incubated with primary antibody (15 µg/mg cell lysate) and matrix (Protein G) and incubated for 2 hour 4°C.Lane 1: Control beadsLane 2: Anti-PLK1 (phospho S137) IP Immunoprecipitation was confirmed by western blot with an HRP-conjugated Mouse monoclonal antibody against pan-PLK1 (1/5000)See Abreview