SMTN Antibody / Smoothelin (FY13280)

Immunofluorescent staining of SMTN using anti-SMTN antibody (red) and anti-Beta Tubulin antibody (green). SMTN was detected in immunocytochemical section of cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-SMTN antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. Cy3 Conjugated Goat Anti-Rabbit IgG and FITC Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of SMTN using anti-SMTN antibody. Lane 1: human Colo320 whole cell lysates, Lane 2: human PC-3 whole cell lysates, Lane 3: human U20S whole cell lysates, Lane 4: human 293T whole cell lysates, Lane 5: rat stomach tissue lysates, Lane 6: rat heart tissue lysates, Lane 7: mouse stomach tissue lysates, Lane 8: mouse heart tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SMTN antibody at 0.25 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. Predicted molecular weight ~99 kDa but commonly observed at ~110 kDa (long form).