Name: | Goat polyclonal antibody to CNP |
Immunogen: | Full length human recombinant protein expressed in and purified from E. coli |
HGNC Name: | CNP |
UniProt: | P09543 |
Molecular Weight: | 46kDa, 48kDa |
Host: | Goat |
Isotype: | |
Species Cross-Reactivity: | Human, rat, mouse, cow, pig |
RRID: | AB_2858266 |
Format: | Affinity purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3 |
Applications: | WB, IF/ICC, IHC |
Recommended Dilutions: | WB: 1:5,000. IF/ICC 1:3,000 |
Storage: | Store at 4°C for short term, for longer term at -20°C |
Goat Polyclonal Antibody to CNP
Cat# GPCA-CNP
$150.00 – $1,000.00
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).
The GPCA-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. The same recombinant protein was used to generate mouse monoclonal anti CNP MCA-1H10, and rabbit and chicken polyclonal anti CNP antibodies, RPCA-CNP and CPCA-CNP. Like GPCA-CNP, these antibodies are excellent markers of myelin and myelinating cells and recognize CNP cleanly on western blots. Mouse select image above left for magnified view.
Chromogenic immunostaining of a formalin fixed paraffin embedded human cerebellum section with goat pAb to CNP, GPCA-CNP, dilution 1:2,000, detected with DAB (brown) using the Vector Elite ABC-HRP detection and reagents with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. The CNP antibody labels myelin axonal sheathes in the granule cell layer and in the molecular layer at the top right. Mouse select image above for larger view.
This antibody was raised against full length recombinant human CNP expressed in and purified from E. coli. The human CNP sequence was derived from NP_149124.3 which was inserted into the eukaryotic expression vector pET30a(+) which adds an N-terminal His tag and some other sequence which is underlined below.
MHHHHHHSSG LVPRGSGMKE TAAAKFERQH MDSPDLGTDD DDKAMADIGS EFMNRGFSRK 60
SHTFLPKIFF RKMSSSGAKD KPELQFPFLQ DEDTVATLLE CKTLFILRGL PGSGKSTLAR 120
VIVDKYRDGT KMVSADAYKI TPGARGAFSE EYKRLDEDLA AYCRRRDIRI LVLDDTNHER 180
ERLEQLFEMA DQYQYQVVLV EPKTAWRLDC AQLKEKNQWQ LSADDLKKLK PGLEKDFLPL 240
YFGWFLTKKS SETLRKAGQV FLEELGNHKA FKKELRQFVP GDEPREKMDL VTYFGKRPPG 300
VLHCTTKFCD YGKAPGAEEY AQQDVLKKSY SKAFTLTISA LFVTPKTTGA RVELSEQQLQ 360
LWPSDVDKLS PTDNLPRGSR AHITLGCAAD VEAVQTGLDL LEILRQEKGG SRGEEVGELS 420
RGKLYSLGNG RWMLTLAKNM EVRAIFTGYY GKGKPVPTQG SRKGGALQSC TII 473
Number of amino acids: 473
Molecular weight: 53286.83
Theoretical pI: 8.80
Amino acid composition:
Ala (A) 33 7.0%
Arg (R) 30 6.3%
Asn (N) 7 1.5%
Asp (D) 32 6.8%
Cys (C) 7 1.5%
Gln (Q) 21 4.4%
Glu (E) 33 7.0%
Gly (G) 39 8.2%
His (H) 12 2.5%
Ile (I) 13 2.7%
Leu (L) 52 11.0%
Lys (K) 42 8.9%
Met (M) 11 2.3%
Phe (F) 22 4.7%
Pro (P) 21 4.4%
Ser (S) 29 6.1%
Thr (T) 28 5.9%
Trp (W) 5 1.1%
Tyr (Y) 14 3.0%
Val (V) 22 4.7%
Total number of negatively charged residues (Asp + Glu): 65
Total number of positively charged residues (Arg + Lys): 72
Ext. coefficient 48735
Abs 0.1% (=1 g/l) 0.915, assuming all pairs of Cys residues form cystines
Ext. coefficient 48360
Abs 0.1% (=1 g/l) 0.908, assuming all Cys residues are reduced
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).
6. Park Y-G, et al. Protection of tissue physicochemical properties using polyfunctional crosslinkers. Nature Biotechnology 10.1038/nbt.4281 doi:10.1038/nbt.428137 (2018).
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