bs-5235R [Primary Antibody]
Smad3 (Ser204) Polyclonal Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: Smad3 Ser204

Specificity: This phosphorylation site is homologous to that of Ser204 in Mouse and Rat.

Modification Site: Ser204

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 4088

Swiss Prot: P84022

Source: KLH conjugated synthetic phosphopeptide derived from human Smad3 around the phosphorylation site of Ser204 [AG(p-S)PN]

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Background:

Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. Has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.

Size: 100ul

Concentration: 1ug/ul

Applications: WB(1:300-5000)
ELISA(1:500-1000)
FCM(1:20-100)
IHC-P(1:200-400)
IHC-F(1:100-500)
IF(IHC-P)(1:50-200)
IF(IHC-F)(1:50-200)
IF(ICC)(1:50-200)
ICC(1:100-500)

Predicted Molecular Weight: 47


Cross Reactive Species: Human
Mouse
Rat

Predicted Cross Reactive Species: Cow
Sheep
Pig
Horse
Rabbit

For research use only. Not intended for diagnostic or therapeutic use.

PRODUCT SPECIFIC PUBLICATIONS
  • Zhang, Hongjun, et al. "Magnolol Attenuates Concanavalin A‐induced Hepatic Fibrosis, Inhibits CD4+ T Helper 17 (Th17) Cell Differentiation and Suppresses Hepatic Stellate Cell Activation: Blockade of Smad3/Smad4 Signalling." Basic & Clinical Pharmacology & Toxicology (2016).Read more>>
  • Huajun Wang. et al. LncRNA NEAT1 promotes proliferation, migration, invasion and epithelial-mesenchymal transition process in TGF-2-stimulated lens epithelial cells through regulating the miR-486-5p/SMAD4 axis. Cancer Cell Int. 2020 Dec;20(1):1-12Read more>>
VALIDATION IMAGES

HUVEC cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with phospho-Smad3 (Ser204) Polyclonal Antibody(bs-5235R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).


Paraformaldehyde-fixed, paraffin embedded Rat uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad3 (Ser204) Polyclonal Antibody, Unconjugated (bs-5235R) at 1:400 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad3 (Ser204) Polyclonal Antibody, Unconjugated (bs-5235R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad3 (Ser204) Polyclonal Antibody, Unconjugated (bs-5235R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human skin cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad3 (Ser204) Polyclonal Antibody, Unconjugated (bs-5235R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human gastric carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad3 (Ser204) Polyclonal Antibody, Unconjugated (bs-5235R) at 1:200 overnight at 4°C, DAB staining.


Lane 1: Mouse Heart lysates; Lane 2: Mouse Kidney lysates; Lane 3: Rat Skin lysates probed with Smad3 Polyclonal Antibody, Unconjugated (bs-5235R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


A431 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad3 (Ser204) Polyclonal Antibody(bs-5235R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).


Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-Smad3 (Ser204)) polyclonal Antibody, Unconjugated (bs-5235R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.