Anti-PLS3 Antibody Picoband®

Western blot analysis of PLS3 using anti-PLS3 antibody (A04134-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human A549 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 antigen affinity purified polyclonal antibody (Catalog # A04134-1) at 0.25 μg/mL overnight at 4°C, 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 is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PLS3 at approximately 71 kDa. The expected band size for PLS3 is at 71,69,66 kDa.

IF analysis of PLS3 using anti-PLS3 antibody (A04134-1).
PLS3 was detected in an immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-PLS3 Antibody (A04134-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of PC-3 cells using anti-PLS3 antibody (A04134-1).
Overlay histogram showing PC-3 cells stained with A04134-1 (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 (A04134-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) 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 (A04134-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human A549 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 antigen affinity purified polyclonal antibody (A04134-1) at 0.25 μg/mL overnight at 4°C, 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 (A04134-1).
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 (2μg) + Hela whole cell lysate (500μg)
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PLS3 antigen affinity purified polyclonal antibody (A04134-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for PLS3 at approximately 71 kDa. The expected band size for PLS3 is at 71 kDa.