The amyloid beta peptide (Ab) of 39 to 43aa (Ab40, Ab42, Ab43) is constitutively produced in brain upon proteolysis of the b-amyloid precursor protein (APP).
In the young and healthy humans, 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 is susceptible to a number of in vivo and in vitro proteases like cathepsin-D and M-13 metalloproteases.
The M-13 family includes several members of zinc-dependent proteases like damage-induced neuronal endopeptidase (DINE), product of phosphate regulating gene with homologies to endopeptidases on the X chromosome (PHEX), neutral endopeptidase 24.11 or neprilysin (NEP) and neprilysin-like proteases (NEPLs).
NEPLs (alpha, -beta, gamma) arise from the alternative splicing of a single NELPS gene and show ~54% sequence homology.
M13 members are generally type II transmembrane proteins consisting of a single polypeptide chain with Zinc binding HEXXH motif, a short cytoplasmic tail, a transmembrane segment and an extra-cytoplasmic domain containing the enzyme active site.
NEPL-g is a 115kD (779 aa) with a Zinc binding HEXXH motif in its extracellular domain.
NEPL-g appears to have a different conformation and substrate specificity than those of NEPL-a.
In comparison to NEPL-a, NEPL-g has an additional sequence of 37aa (311-347) near the center of the polypeptide which possibly inhibits its activity by interfering with proper folding and interaction with the substrate.
Unlike NEP, NEPL-g has no proteolytic activity to either synthetic or Ab peptides and does not appear to be a possible in vivo peptidase for endogenous Ab.
Proteolytic activation of endopeptidase activity of NEPL-g by a partial/complete removal of the unique region (311-347aa) in its sequence has been proposed.
Applications:Suitable for use in ELISA, Western Blotting.
Other applications not tested.
Recommended Dilution:Western Blot: 1:1000-1:5000 for antiserum and 1-10ug/ml for affinity pure using ECL.
ELISA: 1:10,000-1:100,000 using 50-100ng of control peptide/well.
Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
Freeze at -20°C.
Aliquots are stable for at least 12 months at -20°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.
仕様
Size：100ul
Host：rabbit
Source Antibody：mouse
Grade：Serum
Purity：Serum
Form：Supplied as a liquid, neat serum, 40% glycerol.
Specificity：The antigenic/control peptide is conserved in mouse (100%) NEPL-g. No significant sequence homology of is seen with other proteins. Antibody reactivity in various species is not known. The control peptide, because of its low MW (<3kD), is not suitable for Western. It should be used for ELISA or antibody blocking experiments (use 5-10 ug control peptide per 1 ug of aff pure IgG or 1 ul antiserum) to confirm antibody specificity (see below).
Isotype：IgG
Calc Applications Abbrev：E WB
Calc Crossreactivity：Mo
Immunogen：A 15-aa peptide sequence, mapping at the extracellular domain (within the unique 37-aa insert) of mouse NEPL-g (1) was synthesized, coupled to KLH
