Neprilysin, Recombinant, Human, Western Blot Positive Control (Atriopeptidase, Common Acute Lymphocytic Leukemia Antigen, CALLA, Enkephalinase, Neutral Endopeptidase 24.11, NEP, Neutral Endopeptidase, CD10, MME, EPN)

The amyloid b-peptide (Ab) of 39-43aa is constitutively produced in brain upon proteolysis of the b-amyloid precursor protein (APP) and exists as fragments of 40, 42 and 43aa (Ab40, Ab42, Ab43). In the young and healthy humans, the secreted Ab is rapidly catabolized before it can be deposited in the brain. However, upon aging or the onset of familial Alzheimer's disease, alterations in either synthesis or degradation/clearance of Ab may contribute to amyloid depositions in the brain. Ab carries cleavage sites for a number of in vivo and in vitro proteases like cathepsin D and M-13 metalloproteases. The M-13 family comprises several zinc-dependent metalloproteases like DINE, PHEX, KELL, ECE, XCE, neprilysin (NEP) and neprilysin-like proteases (NEPL-a, NEPL-b, NEPL-g).\n\nNEP [variously termed as neutral endopeptidase-24.11 (NEP), neprilysin, enkephalinase, EC3.4.24.11, common acute lymphoblastic leukemia antigen (CALLA), CD10] is the key in vivo enzyme degrading biopeptides Ab, substance-P and enkephalin in brain; atrial natriuretic peptide, bradykinin and endothelin-1 in kidney. It is a 97kD ectoenzyme (mouse, rat, human 750aa) with a large extracellular domain containing its catalytic site, which can degrade Ab on cell surface. NEP is ubiquitous but its high expression in brain is restricted to striatum, olfactory tubercle, substantia nigra, choroids plexes, endopeduncular nucleus, pontine nucleus, and cerebellum and in many peripheral tissues, particularly in brush-border membranes. The poor expression of NEP in the hippocampus and cerebral cortex is reflected in the selective deposition of Ab42 in these regions. NEP is also expressed in soluble form in human plasma and cerebrospinal fluid. NEP can degrade both synthetic and cell secreted Ab40 and Ab42 and may be a good therapeutic target for the treatment of Alzheimer's disease.\n\nSource:\nRecombinant human NEP, expressed in baculovirus cell line Sf21 (Tyr45-Trp743; total of 699 residues).\n\nMolecular Weight:\nPurified protein migrates as ~90kD in SDS-PAGE due to glycosylation. \n\nApplications:\nSuitable for use in Western Blot. It is not suitable for ELISA or other applications where native protein is required. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 10ul/lane. SDS may crystallize in cold conditions. It should redissolve by warming\nbefore taking it from the stock. It should be heated once prior to loading on gels.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20 degrees C. Aliquots are stable for 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

N2030-25

Size

100ul

Applications

ELISA

Form

Supplied as a liquid in SDS-PAGE sample buffer, reduced

Purity

Purified ((same/more than)95%)

References

1. Chen CY et al (1997) J. Immunol. 148, 2817-2825. 2. Malfroy B et al (1987) BBRC 144, 59-66. 3. Bateman RC et al (1989) JBC 264, 6151-6157. 4. Malfroy B et al (1988) J. Exp. Med. 168, 1247-1253. 5. Shipp MA et al (1988) PNAS 85, 4819- 4823. 6. Letarte M et al (1988) J. Exp. Med. 168, 1247-1253. 7. Iwata et al. (2001) Science 292, 1550. 8. Ikeda et al. (2000) J. Biol. Chem. 274, 32469. 9. Boileau et al. (2001) Biochem. J 355, 107. 10. Kiryu-Seo et al. (2000) PNAS 97, 4345.