Phospho-Smad2 (S245/S250/S255) Rabbit mAb for WB and ELISA - Q15796

Core Product Specifications and Parameters

Biological Background: SMAD2 Function and Localization

• SMAD2 (Mothers against decapentaplegic homolog 2) is a receptor-regulated SMAD (R-SMAD) that functions as an intracellular signal transducer and transcriptional modulator for TGF-beta (transforming growth factor) and activin type 1 receptor kinases.
• Upon phosphorylation at its C-terminal SXS motif (serine residues 465 and 467), SMAD2 forms a complex with SMAD4 and translocates to the nucleus where it binds the TGF-beta-responsive element (TRE) in target gene promoters to regulate transcription.
• SMAD2 positively regulates PDPK1 kinase activity by stimulating its dissociation from the inhibitory 14-3-3 protein YWHAQ.
• It has been implicated as a tumor suppressor in colorectal carcinoma. Related references: PMID:8752209
• Subcellularly, SMAD2 shuttles between the cytoplasm and nucleus, consistent with its role in signal transduction.
• The protein is expressed at high levels in skeletal muscle, endothelial cells, heart, and placenta.
• Post-translational modifications include phosphorylation, acetylation, and ubiquitination, which regulate its activity, stability, and interactions.
• Alternative splicing generates multiple isoforms, and the protein contains a zinc ion binding domain (MH1 domain) that mediates DNA binding.

Experimental Guidance and Technical Tips

• This antibody was generated against a phosphorylated peptide; sample lysis buffers should include phosphatase inhibitors (e.g., sodium orthovanadate, sodium fluoride) to maintain phosphorylation status during preparation.
• For Western blotting, consider running both untreated and TGF-β/activin-stimulated cell lysates to confirm activation-dependent band shifts.
• A recommended starting dilution for WB is 1:500–1:2000, but optimal concentration should be determined empirically in your specific sample system.
• In ELISA, using a blocking buffer with low phosphatase activity may reduce background; consider a phospho-peptide competitive assay to verify specificity.
• As a rabbit monoclonal antibody, use an anti-rabbit IgG (H+L) secondary antibody conjugated to HRP or AP for detection.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio partners with diagnostic manufacturers and research institutes to provide a comprehensive portfolio of IVD raw materials, including validated antibody pairs, optimized monoclonal and polyclonal antibodies, and bulk ancillary reagents. For projects targeting the TGF-beta signaling pathway or phospho-SMAD2 detection, CamelBio offers tailored sourcing support to accelerate your assay development from concept to clinic.