ab135637 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 ab135637 at 1/1000 dilution 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-MASTL antibody (ab135637) at 1/100 dilutionLane 1 : HepG2 cell line lysateLane 2 : MDA-MB-231 cell line lysateLane 3 : 293 cell line lysateLane 4 : HeLa cell line lysateLysates/proteins at 35 µg per lane.
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human hepatocarcinoma tissue labelling MASTL with ab135637 at 1/50 dilution. Immunohistochemistry followed by peroxidase conjugation of the secondary antibody and DAB detection.
Flow cytometric analysis of HepG2 cells (bottom histogram) labelling MASTL with ab135637 at 1/10 dilution, compared to a negative control cell (top histogram). FITC-conjugated Goat-anti-Rabbit secondary antibodies were used for the analysis.