ab135585 stained HeLa cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab135585 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
All lanes : Anti-VDAC1/Porin antibody - N-terminal (ab135585) at 1/50 dilutionLane 1 : A375 cell line lysateLane 2 : Mouse heart tissue lysatesLysates/proteins at 35 µg per lane.
Immunohistochemical analysis of VDAC1/Porin in formalin fixed, paraffin embedded Human hepatocarcinoma tissue labelled with ab135585 at a 1/50 dilution, followed by peroxidase conjugation of the secondary antibody and DAB detection.
All lanes : Anti-VDAC1/Porin antibody - N-terminal (ab135585) at 1/50 dilutionLane 1 : 293 cell lysates, non-transfected Lane 2 : 293 cell lysates transiently transfected with VDAC1/Porin Lysates/proteins at 2 µg per lane.
Flow cytometric analysis of VDAC1/Porin in HepG2 cells labelled with ab135585 at a 1/10 dilution (right histogram), compared to a negative control cell (left histogram). FITC-conjugated Goat-anti-Rabbit secondary antibodies were used for the analysis.