bs-3425R
[Primary Antibody]
Smad3 (Ser423 + Ser425) Antibody
DATASHEET
Host: Rabbit
Target Protein: Smad3 Ser423 + Ser425
Specificity: These phosphorylation sites are homologous across the species listed.
Modification Site: Ser423 + Ser425
Clonality: Polyclonal
Isotype: IgG
Entrez Gene: 4088
Swiss Prot: P84022
Source: KLH conjugated synthetic phosphopeptide derived from human Smad3 around the phosphorylation site of Ser423/425 [CS(p-S)V(p-S)]
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
Pig
Predicted Cross Reactive Species:
Dog
Cow
Horse
Chicken
For research use only. Not intended for diagnostic or therapeutic use.
PRODUCT SPECIFIC PUBLICATIONS
- Ghiabi, Pegah, et al. "Breast cancer cells promote a notch-dependent mesenchymal phenotype in endothelial cells participating to a pro-tumoral niche." Journal of translational medicine 13.1 (2015): 27.Read more>>
- Shang, Peijin, et al. "Acetyl-11-Keto-β-Boswellic Acid Attenuates Prooxidant and Profibrotic Mechanisms Involving Transforming Growth Factor-β1, and Improves Vascular Remodeling in Spontaneously Hypertensive Rats." Scientific Reports 6 (2016): 39809.Read more>>
- Bai, Long, et al. "BAMBI promotes porcine granulosa cell steroidogenesis involving TGF-β signaling." Theriogenology (2017).Read more>>
- Zhang, Wen-feng, et al. "Angelica polysaccharides inhibit the growth and promote the apoptosis of U251 glioma cells in vitro and in vivo." Phytomedicine (2017).Read more>>
- Shang et al. Telmisartan improves vascular remodeling through ameliorating prooxidant and profibrotic mechanisms in hypertension via the involvement of transforming growth factor-β1. (2017) Mol.Med.Re. 16:4537-4544Read more>>
- Hu et al. Hydroxysafflor Yellow A Ameliorates Renal Fibrosis by Suppressing TGF-β1-Induced Epithelial-to-Mesenchymal Transition. (2016) PLoS.On. 11:e0153409Read more>>
- Tang et al. Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-κB and TGF-β1/Smad-2/-3 pathways. (2016) Cell.Stress.Chaperones. 21:239-49Read more>>
- Wang et al. Epigallocatechin-3-gallate attenuates unilateral ureteral obstruction-induced renal interstitial fibrosis in mice. (2015) J.Histochem.Cytochem. 63:270-9Read more>>
- Zhang W et al. Smad Anchor for Receptor Activation and Phospho-Smad3 Were Upregulated in Patients with Temporal Lobe Epilepsy.J Mol Neurosci. 2019 May;68(1):91-98.Read more>>
- Wang JQ et al. Knockdown of microRNA-17-5p Enhances the Neuroprotective Effect of Act A/Smads Signal LoopAfter Ischemic Injury. Neurochem Res. 2019 May 15.Read more>>
- Guo LP et al. Smad signaling coincides with epithelial-mesenchymal transition in a rat model of intrauterine adhesion. Am J Transl Res. 2019 Aug 15;11(8):4726-4737. eCollection 2019.Read more>>
- Li L et al. Anti-fibrotic effect of melittin on TRIM47 expression in Human embryonic lung fibroblast through regulating TRIM47 pathway. Life Sci. 2020 Sep 1;256:117893.Read more>>
- Siyu Li. et al. Exploring the liver fibrosis induced by deltamethrin exposure in quails and elucidating the protective mechanism of resveratrol. Ecotox Environ Safe. 2021 Jan;207:111501Read more>>
- Naihua Hu. et al. Forsythiae Fructuse water extract attenuates liver fibrosis via TLR4/MyD88/NF-B and TGF-/smads signaling pathways. J Ethnopharmacol. 2020 Nov;262:113275Read more>>
- Yu Guo. et al. RepSox effectively promotes the induced differentiation of sheep fibroblasts into adipocytes via the inhibition of the TGF?1/Smad pathway. Int J Mol Med. 2021 Aug;48(2):1-13Read more>>
- Haixia Li. et al. Aggravation of hepatic ischemia?reperfusion injury with increased inflammatory cell infiltration is associated with the TGF?/Smad3 signaling pathway. Mol Med Rep. 2021 Aug;24(2):1-1Read more>>
- Xiaoyue Guan. et al. Gremlin aggravates periodontitis via activating the NF-_B signaling pathway. 2022 Jan 06Read more>>
- Guangning Kou. et al. Sesamin Activates Skeletal Muscle FNDC5 Expression and Increases Irisin Secretion via the SIRT1 Signaling Pathway. J AGR FOOD CHEM. 2022;XXXX(XXX):XXX-XXXRead more>>
- Feng Wang. et al. Metformin reduces myogenic contracture and myofibrosis induced by rat knee joint immobilization via AMPK-mediated inhibition of TGF-_1/Smad signaling pathway. CONNECT TISSUE RES. 2022 Jun 2Read more>>
- Kim, Duk Ki. et al. PD-L1-directed PlGF/VEGF blockade synergizes with chemotherapy by targeting CD141+ cancer-associated fibroblasts in pancreatic cancer. NAT COMMUN. 2022 Oct;13(1):1-19Read more>>
- Ping Li. et al. Cyanocobalamin promotes muscle development through the TGF- signaling pathway. FOOD FUNCT. 2022 NovRead more>>
- Xing Li. et al. Inducible nitric oxide synthase (iNOS)-activated Cxcr2 signaling in myeloid cells promotes TGF-dependent squamous cell carcinoma lung metastasis. CANCER LETT. 2023 Aug;570:21633Read more>>
- Genghua Chen. et al. Bulk and single-cell alternative splicing analyses reveal roles of TRA2B in myogenic differentiation. CELL PROLIFERAT. 2023 Sep;:e13545Read more>>
- Demircan Volkan. et al. Evaluation of therapeutic use of a combination of pentoxifylline and vitamin E in radiation-induced renal fibrosis. SCI REP-UK. 2024 Mar;14(1):1-14Read more>>
- Junfeng He. et al. Effect of the TGF-/BMP Signaling Pathway on the Proliferation of Yak Pulmonary Artery Smooth Muscle Cells under Hypoxic Conditions. ANIMALS. 2024 Jan;14(14):2072Read more>>
- Ke Xu. et al. Polystyrene microplastics and di-2-ethylhexyl phthalate co-exposure: Implications for female reproductive health. Environmental Science and Ecotechnology. 2024 Nov;22:100471Read more>>
VALIDATION IMAGES
Lane 1: mouse embryo lysates Lane 2: mouse brain lysates probed with Anti Phospho-Smad3(Ser423/425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band 47kD. Observed band size: 47kD.
Formalin-fixed and paraffin embedded human liver carcinoma labeled with Rabbit Anti-Smad3 (Ser423 + Ser425) Polyclonal Antibody (bs-3425R) at 1:200 followed by conjugation to the secondary antibody and DAB staining\n
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 Smad3 (Ser423 + Ser425) Polyclonal Antibody(bs-3425R)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 Mouse brain; 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse skin; 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat testis; 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat brain; 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat skin; 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat heart; 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) 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 (Ser423 + Ser425) Polyclonal Antibody, Unconjugated (bs-3425R) at 1:200 overnight at 4°C, DAB staining.
Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Heart lysates; Lane 3: Mouse Testis lysates; Lane 4: Mouse Skin lysates; Lane 5: Mouse Kidney lysates; Lane 6: Rat Cerebrum lysates; Lane 7: Rat Testis lysates; Lane 8: Rat Skin lysates; Lane 9: Rat Kidney lysates; Lane 10: Human HUVEC cell lysates; Lane 11: Human A549 cell lysates; Lane 12: Human Hela cell lysates; Lane 13: Human HT1080 cell lysates; Lane 14: Human A431 cell lysates probed with Smad3 Polyclonal Antibody, Unconjugated (bs-3425R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Hela 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 (Ser423 + Ser425) Antibody(bs-3425R)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).