PLS3 Antibody / Plastin 3 (FY12230)

Immunofluorescent staining of PLS3 using anti-PLS3 antibody (green). PLS3 was detected in an immunocytochemical section of cells. 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-PLS3 antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of PLS3 using anti-PLS3 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human Siha whole cell lysates, Lane 4: human PC-3 whole cell lysates, Lane 5: rat stomach tissue lysates, Lane 6: rat kidney tissue lysates, Lane 7: mouse stomach tissue lysates, Lane 8: mouse kidney 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-PLS3 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. A specific band was detected for PLS3 at approximately 71 kDa. The expected band size for PLS3 is at 71,69,66 kDa.

Flow Cytometry analysis of PC-3 cells using anti-PLS3 antibody. Overlay histogram showing PC-3 cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PLS3 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of PLS3 using anti-PLS3 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human SiHa whole cell lysates, Lane 4: human PC-3 whole cell lysates, Lane 5: mouse pancreas 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-PLS3 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-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for PLS3 at approximately 71 kDa. The expected band size for PLS3 is at 71 kDa.

Immunoprecipitating PLS3 in Hela whole cell lysate. Western blot analysis of PLS3 using anti-PLS3 antibody. Lane 1: Hela whole cell lysates (30ug) Lane 2: Rabbit control IgG instead of anti-PLS3 antibody in Hela whole cell lysate. Lane 3: anti-PLS3 antibody (2ug) + Hela whole cell lysate (500ug) After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PLS3 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for PLS3 at approximately 71 kDa. The expected band size for PLS3 is at 71 kDa.