ICC/IF image of ab82541 stained SV40LT-SMC 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 (ab82541, 1µg/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.
All lanes : Anti-smooth muscle Myosin heavy chain 11 antibody (ab82541) at 1 µg/mlLane 1 : PC3 (Human prostate carcinoma cell line) Whole Cell Lysate Whole Cell LysateLane 2 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell LysateLane 3 : Bladder (Human) Tissue Lysate - adult normal tissue (ab44654)Lane 4 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)Lysates/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.
Anti-smooth muscle Myosin heavy chain 11 antibody (ab82541) at 1/2000 dilution + whole cell lysate prepared from human PASMC, P6 at 300 µgSecondaryHRP conjugated goat anti-rabbit polyclonal at 1/2000 dilutiondeveloped using the ECL technique