Analysis Note

ControlOligodendrocyte culture, Brain lysate

Application

This Anti-CNPase Antibody, clone 11-5B is validated for use in IC, IH, IH(P), WB for the detection of CNPase.

Immunohistochemistry:
A previous lot of this antibody was used in IH (10 µg/mL antibody, prepared fresh daily).

Immunocytochemistry:
A previous lot of this antibody was used in IC (10 µg/mL antibody, prepared fresh daily).

Optimal working dilutions must be determined by end user.

General description

The enzyme 2′, 3′-cyclic nucleotide 3′-phosphodi-esterase (CNP) is expressed at high levels by oligodendrocytes in the central nervous system and by Schwann cells in the peripheral nervous system (Sprinkle, 1989). By virtue of this cell-specific expression, CNP is recognized as a characteristic marker for these two myelin-producing glial cell types (Sprinkle, 1989; Kim et al., 1984; Sheedlo & Sprinkle, 1984; McMorris et al., 1984). Beyond its enzymatic activity of cleaving the 2′, 3′-cyclic terminus of nucleotides (Sprinkle, 1989), the physiological role of CNP is still under investigation. CNP activity has been correlated with myelin and myelin formation, and a dramatic decrease in CNP activity is associated with demyelinating diseases such as multiple sclerosis (Sprinkle, 1989). This enzyme is composed of two proteins, CNP1 (46 kD) and CNP2 (48 kD) (Sprinkle, 1989; Sprinkle et al., 1987). Although the ratio of CNP1/CNP2 may vary from species to species, their shared primary sequence is conserved phyIogenetically. CNP has recently been localized to human chromosome 17 by amplification of somatic cell hybrid DNA using the polymerase chain reaction (PCR) and by Southern blotting of Hind Ill genomic DNA digests (Sprinkle et al., 1991). Since anti-CNP reacts with a highly conserved region of the enzyme, it can be considered as pan-anti-CNP (Sprinkle et al., 1987). Anti-CNP can be used as a marker to identify Schwann cells and oligodendrocytes in cell cuIture and in tissue sections, as well as to localize CNP in cell membrane fractions. As CNP is expressed relatively early in postnatal development, anti-CNP is especially useful for the early identification of oligodendrocytes.

Immunogen

Purified human 2', 3'-cyclic nucleotide 3'-phosphodiesterase

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified mouse monoclonal IgG1 in buffer containing 0.02M Phosphate buffer, pH 7.6, 0.25M NaCl with 0.1% sodium azide.

Format: Purified

Quality

Evaluated by Western Blot on Mouse brain lysates.

Western Blotting Analysis:
1:500 dilution of this antibody detected CNPASE 1/2 on 10 µg of Mouse brain lysates.

Specificity

Anti-CNP reacts with both CNP1 and CNP2 in many species.

Target description

48 & 46 kDa