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RNL-3 recognizes an epitope located within the region of amino acids 421-589 of the neuro-endocrine specific protein reticulon-1A (NSP-A), which is also present in the N-terminal part of reticulon-1B (NSP-B). In normal tissues, RNL-3 reacts with brain Purkinje cells, pancreatic islet cells, cells in the pituitary gland and some (peripheral) nerve fibers. In addition, a few epithelia show positive staining.
The antibody may be stored at 4°C. For prolonged storage prepare appropriate aliquots and store at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at 4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance or the concentration of the product.
Recently, a novel gene family has been identified and characterized, designated the Reticulons because the proteins encoded by these genes are anchored to the membranes of the endoplasmic reticulum. Reticulon-1 was formerly designated NSP for Neuroendocrine-Specific-Protein, because it is specifically expressed in neural and neuroendocrine tissues. The NSP-gene has been mapped by fluorescence in situ hybridization to Human chromosome 14q21-q22. The NSP-gene encodes three overlapping proteins, i.e. Reticulon-1A (NSP-A), Reticulon-1B (NSP-B), and Reticulon-1C (NSP-C). These proteins were found to be anchored to membranes of the endoplasmic reticulum through their common carboxy-terminal regions. Reticulon-1A is a protein with a molecular weight (MW) of about 135 kDa, which occurs in various isoforms presumably depending on the degree of phosphorylation of serine residues. In lung cancer diagnosis Reticulon-1A appeared to be a reliable marker for the detection of neuroendocrine differentiation, since most of the small cell lung carcinoma (SCLC) and carcinoid tumors showed expression of Reticulon-1A. Reticulon-1B is a phosphoprotein with a MW of 45 kDa and is restricted to the lung cancer cell line NCI-H82. Reticulon-1B is sofar not found in Human tissues. Reticulon-1C is a protein with a MW of 23 kDa which is not phosphorylated and is found with Reticulon-1A in SCLC (cell lines) and not in non-SCLC (cell cultures).
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