Anti-Phospho-Smad5 (S463/S465) Polyclonal Antibody for WB, IHC-P, ELISA - Q99717

Core Product Specifications and Parameters

Biological Background: Smad5 Function and Localization

• SMAD5 is a member of the SMAD family of intracellular signal transducers and transcriptional modulators, also known as JV5-1, Mothers against decapentaplegic homolog 5, or MADH5.
• Upon BMP ligand binding, SMAD5 is phosphorylated at conserved C-terminal serines (including S463 and S465) by activated type I BMP receptors, then associates with SMAD4 to form a heteromeric complex that translocates to the nucleus to regulate target gene expression. Related references: PMID:9442019, PMID:33510867
• As a transcription factor, phosphorylated SMAD5 recognizes Smad Binding Elements (SBEs) in gene promoters to modulate diverse cellular processes such as embryonic development, cell differentiation, angiogenesis, and tissue homeostasis. Related references: PMID:12064918, PMID:16516194
• In its non-phosphorylated state, SMAD5 operates in the cytoplasm to influence energy metabolism by interacting with hexokinase 1 (HK1) to accelerate glycolysis and by promoting mitochondrial respiration in response to cytoplasmic pH changes. Related references: PMID:28675158
• SMAD5 localizes predominantly to the cytoplasm and nucleus, but also to the mitochondrion, reflecting its multifunctional roles.
• The protein is ubiquitously expressed across tissues and is subject to post-translational modifications including phosphorylation and acetylation; it binds DNA and zinc ions.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide around S463 and S465, suggesting the antibody specifically recognizes Smad5 phosphorylated at these BMP receptor target sites. Consider using phosphatase inhibitors during sample preparation to preserve phosphorylation.
• For Western blot (WB), the predicted molecular weight is 52 kDa; validate detection in relevant BMP-stimulated cell lysates (e.g., human, mouse, rat samples).
• For immunohistochemistry (IHC-P), optimize antigen retrieval and validate in tissue sections where BMP signaling is active; consider using isotype controls to confirm specificity.
• This antibody may cross-react with unphosphorylated Smad5 or other Smad family members; verify specificity by comparing signals in stimulated vs. unstimulated samples or using specific inhibitors.
• In ELISA, use phospho-peptide competition to confirm phospho-specificity, and calibrate using known standards.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is your one-stop partner for IVD raw materials, offering validated antibody pairs, optimized monoclonal/polyclonal antibodies, and bulk ancillary reagents to support diagnostic development from concept to clinic. Our portfolio includes phospho-specific antibodies like this anti-Phospho-Smad5 (S463/S465) pAb, ideal for studying BMP signaling pathways in research and assay development. For rare target raw-material sourcing or customized solutions, our expert team ensures rapid access and technical consultation.