CNPase (2',3'-cyclic Nucleotide 3'-phosphodiesterase) (FITC)

CNPase (2', 3'-cyclic nucleotide 3'-phosphodiesterase [or -phosphohydrolase], EC 3.1.4.37) is present in very high levels in brain and peripheral nerve. This enzyme is found almost exclusively in oligo-dendrocytes and Schwann cells, the cells that form myelin in the central and peripheral nervous system, respectively. CNPase has been very useful as a marker for these particular cell types. Developmentally, this enzyme appears early in oligodendrocytes, earlier than most other myelin proteins. It continues to be expressed at high levels in oligodendrocytes and Schwann cells of adult animals. In various diseases, neurological mutants, and in experimental conditions in which myelin is reduced, CNPase levels may be severely reduced. The enzyme itself exists in two forms in most species (three in mouse) with molecular masses of 48 and 46kD on SDS-PAGE, referred to as CNP2 and CNP1, respectively.Studies continue to elucidate the physiological role of the enzyme. CNPase I (46kD) and CNPase II (48kD) are differentially regulated during development, with the larger protein being expressed earlier than CNPase I during development.||Immunohistochemical localization of CNPase has shown the enzyme to be restricted to oligodendrocytes and Schwann cells. The enzyme appears to be distributed in single and double loose wraps of myelin and not in compact myelin as earlier thought by most investigators. CNPase is located on the inner and outer loops of myelin, paranodally and near the inner surface of the oligodendrocyte membrane. In plaque regions of multiple sclerosis patients, the enzyme is reduced, and when CNS myelin is decreased, CNPase is one of the earlier proteins to be lost from the myelin. In addition, an enzyme that is probably identical to brain CNPase is located in the outer rod segments within the visual system. This protein is also recognized by the CNPase (2',3'-cyclic Nucleotide 3'-phosphodiesterase) Mab Mo xHu (Catalog No: C6205-01). In mixed human gliomas, the enzyme levels are reduced, although about 5% of the oligodendrocytes occasionally show normal positive staining.||Applications:|Suitable for use in Immunocytochemistry, Immunohistochemistry and Western Blot. Other applications have not been tested.||Recommended Dilutions:|Western Blot: 1:1,000 using 10ug of mouse brain lysate as sample. Other suitable samples include myelin, the Wolfgram protein fraction, the SN4 fraction, tissue sections|and mixed glial tumors (oligodendrogliomas, etc.)|Immunohistochemistry: Fresh frozen and paraffin embedded tissue. Note: Recommend microwave pretreatment, ctirate pH 6.0 of paraffin sections.|Immunocytochemistry: Primary oligodendrocyte cultures. |Optimal dilutions to be determined by the researcher.||Storage and Stability:|Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. |Caution: FITC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. ||Note: Applications are based on unconjugated antibody.