HUABIO Smad2 antibody

Smad2
HUABIO
catalog: EM1701-44

mouse monoclonal (C9-B0)
reactivity: human, mouse, rat
application: western blot, immunohistochemistry - paraffin section

Western blot analysis of Smad2 on Hela cell lysate using anti-Smad2 antibody at 1/500 dilution.

ICC staining Smad2 (green) in HUVEC cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

ICC staining Smad2 (green) in A549 cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
quantity: 100μl
price: 360.00 USD
to the supplier

Smad2
HUABIO
catalog: ET1604-22

domestic rabbit monoclonal (SP06-05)
reactivity: human, mouse, rat
application: western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section

Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody (ET1604-22) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HT-29 cell lysate Lane 3: Jurkat cell lysate Lane 4: HL-60 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 52 kDa Observed band size: 58 kDa Exposure time: 1 minute 20 seconds; 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 (ET1604-22) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody (ET1604-22) at 1/1,000 dilution. Lane 1: Jurkat cell lysate (15 µg/Lane) Lane 2: C2C12 cell lysate (15 µg/Lane) Lane 3: Mouse lung tissue lysate (30 µg/Lane) Lane 4: Mouse placenta tissue lysate (30 µg/Lane) Predicted band size: 52 kDa Observed band size: 58 kDa Exposure time: 1 minute 21 seconds; 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 (ET1604-22) 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.

Western blot analysis of Smad2 with anti-Smad2 antibody (ET1604-22) at 1:500 dilution. Lane 1: Wild-type HaCaT whole cell lysate (15 µg). Lane 2: Smad2 knockout HaCaT whole cell lysate (15 µg). ET1604-22 was shown to specifically react with Smad2 in wild-type HaCaT cells. NO band was observed when Smad2 knockout sample was tested. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1604-22, 1:500) was used in 5% BSA at room temperature for 2 hours. Goat anti-Rabbit IgG-HRP antibody at 1:10,000 dilution was used for 1 hour at room temperature.
quantity: 100μl
price: 385.00 USD
to the supplier

Phospho-Smad2 (S250)
HUABIO
catalog: ET1612-32

domestic rabbit monoclonal (SD207-1)
reactivity: human, mouse, rat
application: western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section

Western blot analysis of Phospho-Smad2 (S250) on different lysates with Rabbit anti-Phospho-Smad2 (S250) antibody (ET1612-32) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 200nM PMA for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 55 kDa Exposure time: 30 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 (ET1612-32) at 1/5,000 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.

Western blot analysis of Phospho-Smad2 (S250) on different lysates with Rabbit anti-Phospho-Smad2 (S250) antibody (ET1612-32) at 1/5,000 dilution. Lane 1: C6 cell lysate Lane 2: C6 treated with 200nM PMA for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 55 kDa Exposure time: 2 minutes; 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 (ET1612-32) at 1/5,000 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.

Phospho-Smad2 (S250) was immunoprecipitated from 0.2 mg HeLa treated with 100nM TPA for 30 minutes cell lysate with ET1612-32 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1612-32 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 100nM TPA for 30 minutes cell lysate (input) Lane 2: ET1612-32 IP in HeLa treated with 100nM TPA for 30 minutes cell lysate Lane 3: Rabbit IgG instead of ET1612-32 in HeLa treated with 100nM TPA for 30 minutes cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 35 seconds; ECL: K1802
quantity: 100μl
price: 385.00 USD
to the supplier

Phospho-Smad2 (S255)
HUABIO
catalog: ET1702-34

domestic rabbit monoclonal (JF0882)
reactivity: human, mouse, rat
application: western blot, immunoprecipitation, immunohistochemistry - paraffin section

Western blot analysis of Phospho-Smad2 (S255) on different lysates with Rabbit anti-Phospho-Smad2 (S255) antibody (ET1702-34) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 200nM Calyculin A for 1 hour cell lysate Lane 3: HeLa treated with 200nM Calyculin A for 1 hour cell lysate, then the membrane treated with λpp for 1 hour Lane 4: RAW264.7 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 60 kDa Exposure time: 3 minutes; 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 (ET1702-34) 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 mouse uterus tissue using anti-Phospho-Smad2 (S255) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-34, 1/50) for 30 minutes 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.
quantity: 100μl
price: 385.00 USD
to the supplier

Phospho-SMAD2 (S465+467)
HUABIO
catalog: HA722443

domestic rabbit monoclonal (PSH05-76)
reactivity: human, mouse, rat
application: western blot, immunohistochemistry - paraffin section

Western blot analysis of Phospho-SMAD2 (S465+467) on different lysates with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52/48 kDa Observed band size: 60 kDa Exposure time: 1 minute 20 seconds; ECL: K1802; 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 (HA722443) 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.

Immunocytochemistry analysis of HeLa cells treated with or without 20ng/mL TGF-β1 for 15 minutes labeling Phospho-SMAD2 (S465+467) with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/500 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-SMAD2 (S465+467) antibody (HA722443) at 1/500 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. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunocytochemistry analysis of NIH/3T3 cells treated with or without 20ng/mL TGF-β1 for 15 minutes labeling Phospho-SMAD2 (S465+467) with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/500 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-SMAD2 (S465+467) antibody (HA722443) at 1/500 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. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
quantity: 100μl
price: 385.00 USD
to the supplier

Smad2
HUABIO
catalog: HA750073

domestic rabbit monoclonal (SP06-05)
reactivity: human, mouse, rat
application: western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section

Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody (HA750073) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HT-29 cell lysate Lane 3: Jurkat cell lysate Lane 4: HL-60 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 52 kDa Observed band size: 58 kDa Exposure time: 1 minute 20 seconds; 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 (HA750073) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody (HA750073) at 1/1,000 dilution. Lane 1: Jurkat cell lysate (15 µg/Lane) Lane 2: C2C12 cell lysate (15 µg/Lane) Lane 3: Mouse lung tissue lysate (30 µg/Lane) Lane 4: Mouse placenta tissue lysate (30 µg/Lane) Predicted band size: 52 kDa Observed band size: 58 kDa Exposure time: 1 minute 21 seconds; 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 (HA750073) 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.

Western blot analysis of Smad2 with anti-Smad2 antibody (HA750073) at 1:500 dilution. Lane 1: Wild-type HaCaT whole cell lysate (15 µg). Lane 2: Smad2 knockout HaCaT whole cell lysate (15 µg). HA750073 was shown to specifically react with Smad2 in wild-type HaCaT cells. NO band was observed when Smad2 knockout sample was tested. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (HA750073, 1:500) was used in 5% BSA at room temperature for 2 hours. Goat anti-Rabbit IgG-HRP antibody at 1:10,000 dilution was used for 1 hour at room temperature.
quantity: 100μl
price: 649.00 USD
to the supplier

Phospho-Smad2 (S250)
HUABIO
catalog: HA750291

domestic rabbit monoclonal (SD207-1)
reactivity: human, mouse, rat
application: western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section