The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Predicted molecular weight: 89 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Use at an assay dependent concentration.
ELISA titre using peptide based assay, 1:312500.
RelevanceNeprilysin like Proteases (alpha, beta, gamma) arise from the alternative splicing of a single Neprilysin like Protease gene and show ~54% sequence homology. M13 members are generally type II transmembrane proteins consisting of a single polypeptide chain with Zinc binding HEXXH motif, a short cytoplasmic tail, a transmembrane segment and an extra cytoplasmic domain containing the enzyme active site. Neprilysin like Protease beta is type II transmembrane enzyme containing a single polypeptide chain of 765aa (~110kD) with cytoplasmic and transmembrane domains and a large extracellular C terminal core containing the peptidase active site. In comparison to Neprilysin like Protease alpha, Neprilysin like Protease beta has an additional and unique sequence of 23 aa (41-63 aa) after the TM domain. There are two splice variants of Neprilysin like Protease beta; a secreted isoform of 126kD containing a 23 aa secretion signal sequence and a membrane associated isoform of 110kD. The secreted isoform is much more glycosylated than the membrane isoform.
A number of different sequences for human Neprilysin like Protease beta are reported, with 835, 779, 777, 770, 762 and 753 amino acids in length, having predicted masses of 94.8, 89.4, 89.0, 88.5, 87.2 and 86.2 kDa. The 835 amino acid form has a longer and different carboxytermnius than the other forms. The 753 amino acid form has a 26 residue deletion in the juxtamembrane extracellular region, relative to the other forms, and the 770 form starts at the methionine downstream 10 residues from the other forms. The 762 form uses a different exon in the propeptide domain, and a 17 residue deletion. All of these forms have intact catalytic sites, and are thought to be active enzymes.
Cellular localizationMembrane; Single pass type II membrane protein. Secreted.