EnCor Biotechnology

Chicken Polyclonal Antibody to CNP Cat# CPCA-CNP

$250.00
Description

      The CPCA-CNP antibody was made against the full length recombinant form of human CNP expressed in and purified from E. Coli, and the antibody can be used to identify myelinating cells in cell culture and in sections and to trace axonal projections in sectioned material. We document that the antibody works well not only for western blotting, IF and ICC but also on formalin fixed paraffin embedded sections of human and rodent tissues, select the "Additional Info" for this data. The same recombinant protein was used to generate polyclonal rabbit and goat antibodies to CNP RPCA-CNP and GPCA-CNP and also a mouse monoclonal MCA-1H10. These antibodies are excellent markers of myelin and myelinating cells and recognize CNP cleanly on western blots.

Amount: 100µL of IgY prep
Amount: 100µL of IgY prep
Immunofluorescent analysis of rat cerebellum section stained with chicken pAb to CNP, CPCA-CNP, dilution 1:2,000 in green and costained with rabbit pAb to NF-H, RPCA-NF-H, dilution 1:10,000, in red. Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45μM, and free-floating sections were stained with above antibodies. The CNP antibody stains myelin and oligodendrocytes, cells that create the myelin sheath around axons. The NF-H antibody labels the heavily phosphorylated axonal forms of NF-H which are localized in large projection axons.
Western blot analysis of spinal cord tissue lysates using chicken pAb to CNP, CPCA-CNP, dilution 1:5,000, in red: [1] protein standard (red), [2] mouse, [3] rat, and [4] cow spinal cord. A doublet of bands at 46 and 48kDa correspond to isotypes of the CNP protein. The blot was simultaneously probed with mouse mAb to α-internexin, MCA-2E3, dilution 1:2,000 in green. Major bands in the 64-66 kDa range corresponds to α-internexin.

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Name: CNP, chicken polyclonal, Cat# CPCA-CNP
Immunogen: Full length recombinant human CNP protein
HGNC Name: CNP
UniProt: P09543
Molecular Weight: 46kDa, 48kDa
Host: Chicken
Species Cross-Reactivity: Human, rat, mouse, cow, pig, horse
RRID: AB_2572249
Format: Concentrated IgY preparation plus 0.02% NaN3
Applications: WB, ICC/IF, IHC
Recommended Dilutions: WB: 1:5,000-10,000 IF/IHC: 1:2,000-4,000
Storage: Store at 4°C for short term. For longer term, store at -20°C.

      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. CNP has two isoforms, CNPase 1 (46kDa) and CNPase 2 (48kDa), which are encoded separately by different promoters of the same gene (1). These enzymes are present in very high levels in brain and peripheral nerve, makes up 4% of total CNS myelin protein. They are found almost exclusively in oligodendrocytes and Schwann cells, appearing early in oligodendrocyte development, earlier than most other myelin specific proteins (2). Antibodies to CNP have 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).



Chromogenic Immunostaining of a formalin fixed paraffin embedded human cerebral cortex tissue section with chicken pAb to CNP, CPCA-CNP, dilution 1:4,000, detected in DAB (brown) following the ABC method. Hematoxylin (blue) was used as the counterstain. The CNP antibody stains myelin and oligodendrocytes, cells that produce the myelin sheath around axons. Mouse select image for larger view.

1. Monoh K, Kurihara T, Sakimura K, Takahashi Y. Structure of mouse 2′,3′-cyclic-nucleotide 3′-phosphodiesterase gene. BBRC 165:1213-20 (1989).
2. Kasama-Yoshida H, et al. A comparative study of 2′,3′-cyclic-nucleotide 3′-phosphodiesterase in vertebrates: cDNA cloning and amino acid sequences for chicken and bullfrog enzymes. J. Neurochem. 69:1335–42 (1997).
3. Gravel M, et al. Overexpression of 2′,3′-cyclic nucleotide 3′-phosphodiesterase in transgenic mice alters oligodendrocyte development and produces aberrant myelination. Mol. Cell. Neurosci. 6:453-66 (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 22:547-53 (2001).

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