Anti-Phospho-SMAD1-S463/S465+SMAD5-S463/S465+SMAD9-S465/S467 Rabbit mAb for WB, IHC-P, IF/ICC, ELISA - Q15797/Q99717/O15198

Core Product Specifications and Parameters

Biological Background: SMAD1 Function and Localization

• SMAD1 (Mothers against decapentaplegic homolog 1, also known as BSP1, JV4-1, MADH1) belongs to the intracellular mediator family of SMAD transcription factors.
• Upon BMP ligand binding, SMAD1 is phosphorylated at Ser463/465 by activated type I BMP receptors, then forms a complex with SMAD4 and translocates to the nucleus. Related references: PMID:9335504 PMID:33667543
• Once nuclear, the SMAD1/SMAD4 complex binds Smad Binding Elements (SBEs) to regulate transcription of target genes involved in embryonic development, cell differentiation, and tissue homeostasis.
• SMAD1 mediates degradation of the CREBBP/EP300 repressor SNIP1 through an OAZ1/PSMB4 complex, which further modulates gene expression (By similarity).
• Subcellularly, SMAD1 resides in both the cytoplasm and nucleus, with nuclear translocation dependent on BMP-induced phosphorylation.
• Expression is ubiquitous, with highest levels observed in heart and skeletal muscle.
• Post-translational modifications include phosphorylation (activation), acetylation, and ubiquitination; the protein also features zinc-binding motifs.
• Keywords from UniProt: 3D-structure, Acetylation, Alternative splicing, DNA-binding, Metal-binding, Phosphoprotein, Transcription regulation, Ubl conjugation, Zinc.

Experimental Guidance and Technical Tips

• The antibody was raised against a synthetic phospho-peptide encompassing the conserved phosphorylation motif (SSVS) in SMAD1, SMAD5, and SMAD9; consider confirming specificity by comparing BMP-treated vs. untreated cell lysates in Western blotting.
• For Western blotting, load appropriately treated lysates and include phosphatase inhibitors during preparation; the expected band size is approximately 52 kDa, but phosphorylation may cause slight shifts.
• For IHC-P and IF/ICC, validate the antibody with positive and negative control tissues/cells where BMP signaling is known to be active or inactive; nuclear staining indicates activated SMAD complexes.
• In ELISA, titrate the antibody against phosphorylated vs. non-phosphorylated peptides to verify phospho-specificity.
• Cross-reactivity is reported for human, mouse, and rat; ensure species-specific sequence alignment around the phosphorylation sites before use with other species.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio serves as your one-stop sourcing bridge for IVD raw materials, offering validated phospho-specific antibodies, bulk supplies, and ancillary reagents. We support BMP signaling pathway projects with monoclonal antibodies like this anti-phospho-SMAD1/5/9 and can assist in sourcing rare-target raw materials for diagnostic development. Let our technical expertise help accelerate your research from concept to commercialization.