bsm-52205R [Primary Antibody]
SMAD3 Monoclonal Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: SMAD3

Modification Site: S423/S425

Clonality: Monoclonal

Isotype: IgG

Entrez Gene: 4088

Swiss Prot: P84022

Source: KLH conjugated Synthesised 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 4C. Store at -20C 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

Cross Reactive Species: Human
Mouse
Rat

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

PRODUCT SPECIFIC PUBLICATIONS
  • Xiaoliang Zhou. et al. Ursolic acid inhibits human dermal fibroblasts hyperproliferation, migration, and collagen deposition induced by TGF- via regulating the Smad2/3 pathway. GENE. 2023 May;867:147367Read more>>
VALIDATION IMAGES

Western blot analysis of Phospho-Smad3(S423/S425) on different lysates using anti-Phospho-Smad3(S423/S425) antibody at 1:1,000 dilution. Positive control: Lane 1: A431 cell lysates-treated with TGF β Lane 2: A431 cell lysates-untreated


A431 were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS and stained with Smad3(S423/S425) (5C5) Monoclonal Antibody (bsm-52205R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.


Paraformaldehyde-fixed, paraffin embedded mouse skin section; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37°C for 20min; Antibody incubation with Smad3(S423/S425) (5C5 ) Monoclonal Antibody (bsm-52205R) at 1:50 overnight at 4°C, followed by a conjugated secondary and DAB staining.


Lane 1: A549 cell lysates probed with Smad3(S423/S425) (5C5) Monoclonal Antibody, Unconjugated (bsm-52205R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


Lane 1: Siha cell lysates probed with Smad3(S423/S425) (5C5) Monoclonal Antibody, Unconjugated (bsm-52205R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


Lane 1: Hela cell lysates probed with Smad3(S423/S425) (5C5) Monoclonal Antibody, Unconjugated (bsm-52205R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


Lane 1: Mouse Lung lysates; Lane 2: A549 cell lysates; Lane 3: Hela cell lysates probed with Phospho-Smad3 (Ser423 + Ser425) Monoclonal Antibody, Unconjugated (bsm-52205R ) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.