Anti-Notch1 Polyclonal Antibody for WB, IHC-P, IF/ICC, ELISA - P46531

Core Product Specifications and Parameters

Biological Background: Notch1 Function and Localization

• Notch1 (neurogenic locus notch homolog protein 1) is a single-pass type I transmembrane protein also known as translocation-associated notch protein TAN-1, encoded by the NOTCH1 gene in humans.
• Functions as a receptor for membrane-bound ligands Jagged-1 (JAG1), Jagged-2 (JAG2) and Delta-1 (DLL1), playing a central role in cell-fate determination, differentiation, proliferation, and apoptosis.
• Upon ligand activation, the notch intracellular domain (NICD) is released and forms a transcriptional activator complex with RBPJ/RBPSUH to regulate downstream target genes, including those of the enhancer of split locus.
• Involved in multiple developmental processes: angiogenesis (negatively regulating endothelial cell proliferation and migration), thymic maturation of CD4+ and CD8+ cells, follicular differentiation, cerebellar development via DNER, and left/right symmetry determination through cilia balance at the left-right organiser.
• Represses neuronal and myogenic differentiation; may enhance HIF1A function by sequestering HIF1AN; required for THBS4-mediated protective astrogenesis after injury.
• Subcellular localization includes the cell membrane, late endosome membrane, and nucleus, consistent with its signaling and transcriptional roles.
• Expressed in fetal spleen, brain stem, and lung; in adults, predominantly found in lymphoid tissues.
• Post-translational modifications include glycosylation, disulfide bond formation, hydroxylation, and phosphorylation; the protein is subject to proteolytic processing and ubiquitination.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 2456-2555 of human Notch1 (NP_060087.3), located in the cytoplasmic domain. This region is conserved across human, mouse, and rat, supporting the reported cross-reactivity.
• For Western blotting, a high-molecular-weight standard and a low-percentage (e.g., 6–8%) polyacrylamide gel are recommended due to the large protein size (273 kDa). Prolonged transfer times may be necessary for efficient membrane transfer.
• Validate the antibody in a suitable sample system such as HeLa, Jurkat, or other cell lines known to express Notch1. A positive control tissue like spleen or brain stem may be appropriate for IHC-P.
• For immunohistochemistry (paraffin), antigen retrieval using heat-mediated citrate buffer (pH 6.0) or EDTA buffer (pH 9.0) can be considered, though optimal conditions should be determined experimentally.
• In IF/ICC, permeabilization with Triton X-100 or saponin is suggested to detect intracellular epitopes, as the immunogen is within the cytoplasmic tail.
• Notch1 undergoes proteolytic processing; multiple bands may be observed in WB. The full-length precursor (~300 kDa) and the processed transmembrane/cytoplasmic forms (~120 kDa and ~110 kDa) can be detected, depending on the cell type and activation state.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, laboratories, and research institutes with a comprehensive one-stop sourcing platform for IVD raw materials. For Notch1-related research and assay development, we supply validated antibody pairs, optimized polyclonal and monoclonal antibodies, bulk ancillary reagents, and access to rare-target raw materials. Our integrated supply chain and technical support streamline your project from concept to clinical application, ensuring you have the critical reagents needed for developmental biology and Notch signaling studies.