Smad3 [p Ser423, p Ser425] Antibody (ST0493)

Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of Phospho-Smad3(S423/S425) on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody ( 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.

Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Lane 1: K562 cells, whole cell lysate, 10ug/lane, Lane 2: K562 cells treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane. All lanes : Anti-Phospho-Smad3(S423/S425) antibody at 1:500 dilution. Anti-GAPDH antibody at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution. Predicted band size: 48 kDa Observed band size: 55 kDa Blocking and diluting buffer: 5% BSA. Exposure time: 3 minutes 43 seconds

Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of Smad3 [p Ser423, p Ser425] on different lysates with Rabbit anti-Smad3 [p Ser423, p Ser425] antibody at 1/500 dilution. Lane 1: A549 cell lysate Lane 2: K562 cell lysate Lysates/proteins at 10 ug/Lane. Predicted band size: 48 kDa Observed band size: 60 kDa Exposure time: 2 minutes; 10% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry-Paraffin: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of paraffin-embedded mouse skin tissue using anti-Phospho-Smad3(S423/S425) antibody. Counter stained with hematoxylin.

Immunohistochemistry-Paraffin: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of paraffin-embedded human large intestine tissue with Rabbit anti-Phospho-Smad3 [p Ser423, p Ser425] antibody washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemistry-Paraffin: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of paraffin-embedded mouse skin tissue with Rabbit anti-Smad3 [p Ser423, p Ser425] antibody washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513]

Western blot analysis on different lysates with NBP2-67513 at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: A549 whole cell lysate Lane 2: A549 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate Lane 3: C2C12 whole cell lysate Lane 4: C2C12 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 48 kDa Observed band size: 55 kDa Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. NBP2-67513 at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG [HRP] Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513]

Western blot analysis on different lysates with NBP2-67513 at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa starved overnight then treated with 10ng/mL TGF-beta 1 for 30 minutes cell lysate Lane 3: A549 cell lysate Lane 4: A549 treated with 5ng/mL TGF-beta1 for 24 hours cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: NIH/3T3 starved overnight then treated with 10ng/mL TGF-beta 1 for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 48 kDa Observed band size: 55 kDa Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. NBP2-67513 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG [HRP] Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513]

Western blot analysis of HeLa lysates with NBP2-67513 at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 10 ng/ml TGF beta1 for 30 minutes cell lysate Lane 3: HeLa treated with 10 ng/ml TGF beta1 for 30 minutes cell lysate，then the membrane treated with lambda pp for 1 hour Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: NBP2-67513, 1/2,000 in primary antibody dilution buffer, overnight at 4 ℃ Secondary antibody: Goat Anti-Rabbit IgG [HRP] Secondary Antibody, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 48 kDa Observed band size: 48 kDa