Mixed neuron-glial cell cultures stained with CPCA-CNP (red) and our monoclonal antibody against GFAP (green, MCA-5C10). The CNP antibody stains strongly in oligodendrocytes, whereas GFAP labels the intermediate filament in astrocytes. Blue is DNA staining showing the nuclei of these and numerous other cells, most of which are neurons and which do not express either CNP or GFAP.
|Blot of rat brain tissue homogenates blotted with CPCA-CNP at 1:5,000 using LiCor infrared imager. CPCA-CNP binds strongly and specifically to CNP at ~48 kDa. Numbers at left show positions of protein standards of indicated molecular weight in kDa.
||Antibody to 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP)
||Chicken Polyclonal antibody to CNP
||Full length human recombinant protein
||Antibody is supplied as IgY prep
||Human, rat, mouse
||Western blot, ICC/IF, IHC
|Suggestions for use
||Shipped on ice. Store at 4°C. For long term storage, leave frozen at -20°C. Avoid freeze / thaw cycles.
The 2′, 3′-Cyclic Nucleotide 3′-Phosphodiesterase (CNP), is an enzyme which catalyzes the hydrolysis of 2′, 3′-cyclic nucleotides to 2′-nucleotides, These cyclic nucleotides are structurally different from the better known and studied 3′-5′-cyclic nucleotides of which the best known example is cyclic AMP. The CNP enzyme hydrolyzes 2′,3′-cyclic nucleosidephosphates to produce nucleoside 2′-phosphates and water. CNP has two isoforms, CNPase 1 (46kDa) and CNPase 2 (48kDa), which are encoded separately by different promoters of the same gene (1). This enzyme is present in very high levels in brain and peripheral nerve, makes up 4% of total CNS myelin protein. It is found almost exclusively in oligodendrocytes and Schwann cells. It appears early in oligodendrocyte development, earlier than most other myelin proteins and continues to be expressed at high levels in these cells of adult animals (2). Antibody to CNP has been very useful as a marker for these particular cell types. CNP is thought to play a critical role in the events leading up to myelination, for the oligodendrocytes overexpressing CNP appear to mature earlier in development, resulting in earlier maximum gene expression for myelin basic proteins (3). It has been reported that CNP is also associated with microtubules in brain tissue and may promote microtubule assembly. CNP can link tubulin to cellular membranes, and may regulate cytoplasmic microtubule distribution (4). In various diseases, neurological mutants, and in experimental conditions in which myelin is reduced, CNP levels may also be severely reduced. Decreased brain levels of CNP have also been reported in Down syndrome and Alzheimer’s disease (5).
This antibody was raised against full length recombinant human CNP expressed in and purified from E. coli. The HGNC name for this protein is CNP.
1. Monoh K, Kurihara T, Sakimura K, Takahashi Y. Structure of mouse 2′,3′-cyclic-nucleotide 3′-phosphodiesterase gene. Biochem Biophys Res Commun 165:1213-1220 (1989).
2. Kasama-Yoshida H, Tohyama Y, Kurihara T, Sakuma M, Kojima H, Tamai Y. A comparative study of 2′,3′-cyclic-nucleotide 3′-phosphodiesterase in vertebrates: cDNA cloning and amino acid sequences for chicken and bullfrog enzymes. Journal of Neurochemistry 69:1335–42 (1997).
3. Gravel M, Peterson J, Yong VW, Kottis V, Trapp B, Braun PE. Overexpression of 2′,3′-cyclic nucleotide 3′-phosphodiesterase in transgenic mice alters oligodendrocyte development and produces aberrant myelination. Molecular and Cellular Neurosciences t:453-66 (1996).
4. Bifulco M, Laezza C, Stingo S, Wolff J. 2′,3′-Cyclic nucleotide 3′-phosphodiesterase: a membrane-bound, microtubule-associated protein and membrane anchor for tubulin. PNAS 99:1807–11 (2001).
5. Vlkolinský R, Cairns N, Fountoulakis M, Lubec G.Decreased brain levels of 2′,3′-cyclic nucleotide-3′-phosphodiesterase in Down syndrome and Alzheimer’s disease. Neurobiol Aging 22:547-553 (2001).
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