bs-5459R
[Primary Antibody]
Smad3 (Ser213) Polyclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: Smad3 Ser213
Modification Site: Ser213
Clonality: Polyclonal
Isotype: IgG
Entrez Gene: 4088
Source: KLH conjugated synthetic phosphopeptide derived from human Smad3 around the phosphorylation site of Ser213 [PM(p-S)PA]
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:
Smad3 is a 50 kDa member of a family of proteins that act as key mediators of TGF beta superfamily signaling in cell proliferation, differentiation and development. The Smad family is divided into three subclasses: receptor regulated Smads, activin/TGF beta receptor regulated (Smad2 and 3) or BMP receptor regulated (Smad 1, 5, and 8); the common partner, (Smad4) that functions via its interaction to the various Smads; and the inhibitory Smads, (Smad6 and 7). Activated Smad3 oligomerizes with Smad4 upon TGF beta stimulation and translocates as a complex into the nucleus, allowing its binding to DNA and transcription factors. Phosphorylation of the two TGF beta dependent serines 423 and 425 in the C terminus of Smad3 is critical for Smad3 transcriptional activity and TGF beta signaling.
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
Chicken
Rabbit
Guinea Pig
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>>
- Gao, Lili, et al. "Glycyrrhizic acid alleviates bleomycin-induced pulmonary fibrosis in rats." Frontiers in pharmacology 6 (2015).Read more>>
- Zhou et al. Induced pluripotent stem cell-conditioned medium suppresses pulmonary fibroblast-to-myofibroblast differentiation via the inhibition of TGF-β1/Smad pathway. (2018) Int.J.Mol.Med. 41:473-484Read more>>
- S Q et al. Cyanidin-3-glucoside from black rice prevents renal dysfunction and renal fibrosis in streptozotocin-diabetic rats. Journal of Functional Foods,(2020)72, 104062.Read 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 Smad3 (Ser213) Polyclonal Antibody(bs-5459R)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 Human breast 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 (Ser213) Polyclonal Antibody, Unconjugated (bs-5459R) at 1:400 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human brain glioma; 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 (Ser213) Polyclonal Antibody, Unconjugated (bs-5459R) at 1:400 overnight at 4°C, DAB staining.
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 (Ser213) Polyclonal Antibody, Unconjugated (bs-5459R) 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 (Ser213) Polyclonal Antibody, Unconjugated (bs-5459R) 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 (Ser213) Polyclonal Antibody, Unconjugated (bs-5459R) 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 (Ser213) Polyclonal Antibody, Unconjugated (bs-5459R) 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 (Ser213) Polyclonal Antibody, Unconjugated (bs-5459R) at 1:200 overnight at 4°C, DAB staining.
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 (Ser213) Polyclonal Antibody(bs-5459R)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).