Phospho-Smad5 (S463/S465) Rabbit Monoclonal Antibody for WB, ELISA - Q99717

Core Product Specifications and Parameters

Biological Background: Smad5 Function and Localization

• SMAD5 (Mothers against decapentaplegic homolog 5, also known as JV5-1, MADH5) is a transcriptional regulator belonging to the SMAD family. It plays essential roles in embryonic development, cell differentiation, angiogenesis, and tissue homeostasis. Related references: PMID:12064918 PMID:16516194

• Upon BMP ligand stimulation, SMAD5 is phosphorylated at Ser463 and Ser465 by activated type I BMP receptors. This phosphorylation promotes its association with SMAD4 to form a heteromeric complex that translocates to the nucleus, where it regulates gene expression by binding to Smad Binding Elements (SBEs) in promoter regions. Related references: PMID:9442019 PMID:33510867

• In addition to its transcription factor role, non-phosphorylated SMAD5 functions in the cytoplasm to regulate energy metabolism. It enhances mitochondrial respiration and glycolysis by interacting with hexokinase 1 (HK1), thereby accelerating glycolysis in response to cytoplasmic pH changes. Related references: PMID:28675158

• The protein localizes to the cytoplasm, nucleus, and mitochondrion, reflecting its diverse roles in signaling and metabolism.

• SMAD5 is ubiquitously expressed, consistent with its widespread involvement in fundamental cellular processes.

• Key post-translational modifications include phosphorylation at S463/S465 (critical for activation) and acetylation. The protein contains zinc-binding domains and has been confirmed by direct protein sequencing.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide spanning the dual phosphorylation site (S463/S465). Consider using non-phosphorylated peptide blocking controls to confirm phosphorylation specificity in Western blot.
• For Western blot, the predicted molecular weight is 52 kDa. Validate the band in relevant human cell lines treated with BMP ligands to induce Smad5 phosphorylation. Non-treated or phosphatase-treated lysates can serve as negative controls.
• In ELISA applications, titrate the antibody to determine the optimal working concentration. Phospho-peptide coated plates may enhance specificity.
• Since SMAD5 is ubiquitous, ensure sample preparation and loading controls are consistent across tissues or cell types.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is a dedicated one-stop sourcing partner for IVD raw materials, supporting diagnostic manufacturers and research laboratories from concept to clinic. For BMP signaling and Smad5 research, we offer validated antibody pairs, optimized monoclonal/polyclonal antibodies, bulk ancillary reagents, and can assist with rare-target raw-material sourcing. Whether you need off-the-shelf reagents or custom solutions, our platform streamlines your procurement and accelerates assay development.