CST, 8063S, Phospho-NEDD4L (Ser448) Antibody

Polyclonal Antibody for studying NEDD4L (Ser448) phosphate. Validated for Western Blotting,Immunoprecipitation. Highly specific and rigorously validated in-house, Phospho-NEDD4L (Ser448) Antibody (CST #8063) is ready to ship. Product Usage Information Western Blotting: 1:1000 Immunoprecipitation: 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation Specificity / Sensitivity Phospho-NEDD4L (Ser448) Antibody detects endogenous levels of NEDD4L protein only when phosphorylated at Ser448. Species Reactivity: Human Source / Purification Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser448 of human NEDD4L protein (corresponds to mouse Ser477 or rat Ser436). Antibodies are purified by protein A and peptide affinity chromatography. Background Neural precursor expressed, developmentally down-regulated protein 4 (NEDD4) was originally identified as a gene that is highly expressed in the early mouse embryonic central nervous system (1). Subsequently, a family of NEDD4-like proteins have been defined that includes seven members in humans (2). NEDD4 and NEDD4-like (NEDD4L) proteins contain multiple functional domains including a calcium-dependent phospholipid and membrane binding domain (C2 domain), two to four protein binding domains (WW domains), and an E3 ubiquitin-protein ligase domain (HECT domain). NEDD4 and NEDD4L have been shown to downregulate both neuronal voltage-gated Na channels (NaVs) and epithelial Na channels (ENaCs) in response to increased intracellular Na concentrations (3,4). The WW domains of NEDD4 bind to PY motifs (amino acid sequence PPXY) found in multiple NaV and ENaC proteins; ubiquitination of these proteins is mediated by the HECT domain of NEDD4 and results in their internalization and removal from the plasma membrane. Research studies have shown that mutation of the PY motifs in ENaC proteins is associated with Liddle's syndrome, an autosomal dominant form of hypertension (5). In addition to targeting sodium channels, NEDD4L has also been shown to negatively regulate TGF-β signaling by targeting Smad2 for degradation (6). Mouse and human NEDD4 are rapidly cleaved by caspase proteins during apoptosis, although the significance of this cleavage is not clear (7). NEDD4L can be phosphorylated at Ser342 and Ser448 by several AGC kinase family members including SGK1, Akt, and PKA (8-11). Phosphorylation at these sites inhibits NEDD4L function as a suppressor of ENaCs and as a regulator of TGF-β signaling through its effects on Smad2/3 binding (8-11). Alternate Names E3 ubiquitin-protein ligase NEDD4-like; FLJ33870; HECT-type E3 ubiquitin transferase NED4L; hNedd4-2; KIAA0439; NED4L; NEDD4 like E3 ubiquitin protein ligase; Nedd4-2; NEDD4.2; NEDD4L; NEDL3; neural precursor cell expressed, developmentally down-regulated 4-like; neural precursor cell expressed, developmentally down-regulated 4-like, E3 ubiquitin protein ligase; neural precursor cell expressed, developmentally down-regulated gene 4-like; PVNH7; RSP5; ubiquitin-protein ligase NEDD4-like; ubiquitin-protein ligase Rsp5 Specification REACTIVITY: H SENSITIVITY: Endogenous MW (kDa): 110, 135 SOURCE: Rabbit