Phospho-Smad3 (S423 + S425) | HUABIO HA750172 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

Phospho-Smad3 (S423 + S425)

catalog :

HA750172

quantity :

100μl

price :

649.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

ST0493

reactivity :

human, mouse

application :

western blot, immunohistochemistry - paraffin section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of Phospho-Smad3 (S423 + S425) on different lysates with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) 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; 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 (HA750172) 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 (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

image 2 :

Western blot analysis of Phospho-Smad3(S423/S425) on K562 cell lysates. 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 (HA750172) at 1/2,000 dilution. Anti-GAPDH antibody (ET1601-4) 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

image 3 :

Western blot analysis of Phospho-Smad3 (S423 + S425) on different lysates with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) at 1/2,000 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: 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-β1 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; 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 (HA750172) at 1/2,000 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.

product information

SKU :

HA750172

Target name :

Phospho-Smad3 (S423 + S425)

Species reactivity :

Human,Mouse

Applications :

WB,IF-Cell,IF-Tissue,IHC-P

Conjugate :

Non-conjugated

Immunogen :

Synthetic phospho-peptide corresponding to residues surrounding Ser423 and 425 of human Smad3.

Uniprot id :

P84022>SwissProt: P84022 Human;SwissProt: Q8BUN5 Mouse

Host :

Rabbit

Clone number :

ST0493

Isotype :

IgG

Size :

100μl

List Price :

649.00 USD

Storage Buffer :

PBS (pH7.4).

Form :

Liquid

Storage Instruction :

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Purity :

Protein A affinity purified.

Product type :

Recombinant Rabbit monoclonal Antibody

Positive control :

A549 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate, C2C12 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate, HeLa starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate, NIH/3T3 starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate, A549, human large intestine tissue, mouse skin tissue, mouse kidney tissue.

Molecular wt :

Predicted band size: 48 kDa

Subcellular location :

Cytoplasm, Nucleus.

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:2,000-1:5,000 ;IF-Cell: 1:500 ;IF-Tissue: 1:200 ;IHC-P: 1:200-1:1,000

Advanced Validation :

Cell treatment (CT)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/HA750172_4.jpg

Pic legend4 :

Immunocytochemistry analysis of A549 cells labeling Phospho-Smad3 (S423 + S425) with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) 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-Smad3 (S423 + S425) antibody (HA750172) 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.

Pic img5 :

https://storage.huabio.cn/huabio/productImg/HA750172_5.jpg

Pic legend5 :

Immunohistochemical analysis of paraffin-embedded human large intestine tissue with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) at 1/200 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 (HA750172) 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.

Pic img6 :

https://storage.huabio.cn/huabio/productImg/HA750172_6.jpg

Pic legend6 :

Immunohistochemical analysis of paraffin-embedded mouse skin tissue with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) at 1/500 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 (HA750172) 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.

Pic img7 :

https://storage.huabio.cn/huabio/productImg/HA750172_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) 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 (HA750172) 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.

Pic img8 :

https://storage.huabio.cn/huabio/productImg/HA750172_8.jpg

Pic legend8 :

Immunofluorescence analysis of paraffin-embedded human large intestine tissue labeling Phospho-Smad3 (S423 + S425) with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) at 1/200 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750172, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Pic img9 :

https://storage.huabio.cn/huabio/productImg/HA750172_9.jpg

Pic legend9 :

Immunofluorescence analysis of paraffin-embedded mouse kidney tissue labeling Phospho-Smad3 (S423 + S425) with Rabbit anti-Phospho-Smad3 (S423 + S425) antibody (HA750172) at 1/200 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750172, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

more info or order :

HUABIO product webpage