Phospho-eIF4EBP1 (S65) Rabbit Monoclonal Antibody for WB, ELISA - Q13541

Core Product Specifications and Parameters

Biological Background: eIF4EBP1 Function and Localization

• eIF4EBP1 (eukaryotic translation initiation factor 4E-binding protein 1), also known as PHAS-I, functions as a key repressor of cap-dependent translation initiation by binding to the eukaryotic initiation factor 4E (eIF4E).
• In its hypophosphorylated state, eIF4EBP1 competes with eIF4G (eIF4G1 and eIF4G3) for binding to eIF4E, thereby preventing assembly of the eIF4F complex and inhibiting translation initiation.
• Hyperphosphorylation of eIF4EBP1, often mediated by the mTORC1 pathway, induces its dissociation from eIF4E, allowing eIF4F complex formation and translational activation.
• eIF4EBP1 integrates signals from hormones, growth factors, and nutrients via the MAP kinase and mTOR signaling pathways to control protein synthesis and cell growth.
• It is a phosphoprotein with multiple phosphorylation sites, including Ser65, that modulate its affinity for eIF4E and its ability to regulate translation.
• Subcellularly, eIF4EBP1 localizes to both the cytoplasm and the nucleus, consistent with its roles in translational control and possibly nuclear-cytoplasmic shuttling.
• The protein is post-translationally modified by acetylation and ubiquitin-like conjugation (Ubl) in addition to phosphorylation, highlighting complex regulatory mechanisms.

Experimental Guidance and Technical Tips

• Because this antibody targets a specific phosphorylation site (Ser65), confirm the specificity of detection by treating cell lysates with lambda phosphatase or by comparing signals from wild-type versus phospho-deficient mutants.
• For Western blot, use standard procedures but ensure that lysis buffers contain phosphatase inhibitors to preserve phosphorylation states. A blocking agent like BSA may help reduce nonspecific binding typical of phospho-specific antibodies.
• In ELISA, consider coating with the peptide immunogen as a positive control and include a non-phosphorylated peptide to verify phospho-specificity.
• Activation of mTORC1 (e.g., by insulin or serum stimulation) increases Ser65 phosphorylation, while inhibition (e.g., rapamycin) decreases it; include appropriate stimulatory or inhibitory controls in your experiments.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers and research teams with a comprehensive selection of IVD raw materials, including validated antibody pairs, optimized monoclonal and polyclonal antibodies, and bulk ancillary reagents. For projects targeting eIF4EBP1 and translation regulation, we offer phospho-specific detection tools and can source rare or custom raw materials to meet your development needs. Partner with us to bridge the gap from concept to clinic with reliable, high-quality reagents.