THYN1 Antibody / Thymocyte nuclear protein 1 (FY12881)

Immunofluorescent staining of THYN1 using anti-THYN1 antibody (green) and anti-Alpha Tubulin antibody (red). THYN1 was detected in an immunocytochemical section of U2OS 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-THYN1 antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG and Cy3 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 THYN1 using anti-THYN1 antibody. Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human LNCAP whole cell lysates, Lane 2: human U2OS whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human HepG2 whole cell 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-THYN1 antibody at 0.5 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 an ECL Plus Western Blotting Substrate. A specific band was detected for THYN1 at approximately 26 kDa. The expected molecular weight of THYN1 is ~26 kDa.

Immunoprecipitating THYN1 in Hela whole cell lysate. Western blot analysis of THYN1 using anti-THYN1 antibody. Lane 1: Hela whole cell lysates (30ug), Lane 2: Rabbit control IgG instead of anti-THYN1 antibody in Hela whole cell lysate, Lane 3: anti-THYN1 antibody (2ug) + Hela whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-THYN1 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 THYN1 at approximately 26 kDa. The expected molecular weight of THYN1 is at 26 kDa.

Flow Cytometry analysis of HepG2 cells using anti-THYN1 antibody. Overlay histogram showing HepG2 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-THYN1 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.