Phospho-Smad5 (S463 + S465)
HUABIO
catalog: ET1605-5
domestic rabbit monoclonal (SY09-03)
reactivity: human, mouse, rat
application: western blot, immunohistochemistry - paraffin section

Western blot analysis of Phospho-Smad5 (S463 + S465) on different lysates with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (ET1605-5) at 1/1,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 20 ng/mL TGFβ1 for 15 minutes whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 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. The primary antibody (ET1605-5) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Phospho-Smad5 (S463 + S465) on different lysates with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (ET1605-5) at 1/500 dilution. Lane 1: Rat brain tissue lysate Lane 2: Mouse brain tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: 2 minutes; 8% 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 (ET1605-5) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of HeLa cells treated with 20ng/mL TGFβ1 for 15 minutes labeling Phospho-Smad5 (S463 + S465) with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (ET1605-5) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (ET1605-5) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
quantity: 100μl
price: 385.00 USD
to the supplier
Phospho-Smad1/5/9 (S463/S465/S467)
HUABIO
catalog: HA722566
domestic rabbit monoclonal (JE59-46)
reactivity: human, mouse, rat
application: western blot, immunohistochemistry

Western blot analysis of Phospho-Smad1/5/9 (S463/S465/S467) on different lysates with Rabbit anti-Phospho-Smad1/5/9 (S463/S465/S467) antibody (HA722566) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 10ng/mL BMP4 for 1 hour cell lysate Lane 3: HeLa treated with 10ng/mL BMP4 for 1 hour cell lysate, then the membrane treated with λpp for 1 hour Lane 4: NIH/3T3 cell lysate Lane 5: NIH/3T3 treated with 10ng/mL BMP4 for 1 hour cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: Lane 1-3: 3 minutes; Lane 4-5: 42 seconds; ECL: K1801; 4-20% 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 (HA722566) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Phospho-Smad1/5/9 (S463/S465/S467) antibody (HA722566) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722566) at 1/1,000 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 analysis of Phospho-Smad1/5/9 (S463/S465/S467) on different lysates with Rabbit anti-Phospho-Smad1/5/9 (S463/S465/S467) antibody (HA722566) at 1/1,000 dilution. Lane 1: C6 cell lysate Lane 2: C6 treated with 10ng/mL BMP4 for 1 hour cell lysate Lane 3: C6 treated with 10ng/mL BMP4 for 1 hour cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 10 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: 1 minute 18 seconds; ECL: K1801; 4-20% 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 (HA722566) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
quantity: 100μl
price: 385.00 USD
to the supplier
Phospho-Smad5 (S463 + S465)
HUABIO
catalog: HA750089
domestic rabbit monoclonal (SY09-03)
reactivity: human, mouse, rat
application: western blot, immunohistochemistry - paraffin section

Western blot analysis of Phospho-Smad5 (S463 + S465) on different lysates with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (HA750089) at 1/1,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 20 ng/mL TGFβ1 for 15 minutes whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 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. The primary antibody (HA750089) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Phospho-Smad5 (S463 + S465) on different lysates with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (HA750089) at 1/500 dilution. Lane 1: Rat brain tissue lysate Lane 2: Mouse brain tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: 2 minutes; 8% 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 (HA750089) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of HeLa cells treated with 20ng/mL TGFβ1 for 15 minutes labeling Phospho-Smad5 (S463 + S465) with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (HA750089) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Smad5 (S463 + S465) antibody (HA750089) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
quantity: 100μl
price: 649.00 USD
to the supplier