Phospho-Tuberin (Ser939) Antibody / TSC2 [clone 32T70] (FY12943)

Immunohistochemical staining of Tuberin/TSC2 (Phospho-S939) using anti-Phospho-Tuberin (Ser939) antibody. Tuberin/TSC2 (Phospho-S939) was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with a dilution of 1:50 rabbit anti-Phospho-Tuberin (Ser939) antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Western blot analysis of Tuberin/TSC2 (Phospho-S939) using anti-Phospho-Tuberin (Ser939) antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human PC-3 whole cell lysates, Lane 2: human SIHA whole cell lysates, Lane 3: human MCF-7 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-Tuberin/TSC2 (Phospho-S939) antibody at a dilution of 1:500 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 band is detected just below the 250 kDa marker, consistent with the known slower migration of hyperphosphorylated TSC2 relative to its ~201 kDa predicted mass. The upper mobility reflects multi-site phosphorylation of TSC2 and compression of large proteins on an 8% gel.

Immunohistochemical staining of Tuberin/TSC2 (Phospho-S939) using anti-Phospho-Tuberin (Ser939) antibody. Tuberin/TSC2 (Phospho-S939) was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with a dilution of 1:50 rabbit anti-Phospho-Tuberin (Ser939) antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Western blot analysis of Tuberin/TSC2 using anti-Tuberin/TSC2 antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human SH-SY5Y whole cell lysates, Lane 2: human SIHA whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human 293T 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-Tuberin/TSC2 antibody at 1:500 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 band is detected just below the 250 kDa marker, consistent with the known slower migration of hyperphosphorylated TSC2 relative to its ~201 kDa predicted mass. The upper mobility reflects multi-site phosphorylation of TSC2 and compression of large proteins on an 8% gel.

Immunohistochemical staining of Tuberin/TSC2 using anti-Tuberin/TSC2 antibody. Tuberin/TSC2 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-Tuberin/TSC2 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Immunohistochemical staining of Tuberin/TSC2 using anti-Tuberin/TSC2 antibody. Tuberin/TSC2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-Tuberin/TSC2 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.