BACE-1 is an aspartic protease and an integral membrane protein (1, 2). It is the major beta secretase, and together with the gamma secretase, is responsible for generating the amyloid beta peptide (A beta ) from the amyloid precursor protein (APP) (3, 4). Because A beta is a major component of amyloid plaques, BACE-1 has been implicated in the onset and/or progression of Alzheimer's disease. High levels of BACE-1 activity are sufficient to elicit neurodegeneration and neurological decline in vivo, indicating that inhibiting BACE-1 may block not only A beta -dependent but also A beta -independent pathogenic mechanisms (5). In addition to APP, BACE-1 also cleaves APP-like proteins 1 and 2, the cell adhesion protein P-selectin glycoprotein ligand-1 and beta -galactoside alpha 2,6-sialyltransferase, implying that BACE-1 may have additional functions involving the ectodomain shedding of membrane proteins (6-8). The purified recombinant mouse BACE-1 corresponds to the ectodomain with the activity as described in Activity Assay Protocol.
Recombinant Mouse BACE-1 Protein, CF
R&D Systems | Catalog # 2976-AS
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse BACE-1 Protein (2976-AS)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse BACE-1 protein
Thr22-Thr457, with a C-terminal 10-His tag
Thr22-Thr457, with a C-terminal 10-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Thr22
Predicted Molecular Mass
50 kDa
SDS-PAGE
68 kDa, reducing conditions
Activity
Measured by its ability to cleave a fluorogenic peptide substrate, Mca-SEVNLDAEFRK(Dpn)RR-NH2 (Catalog # ES004).
The specific activity is >2 pmol/min/µg, as measured under the described conditions.
The specific activity is >2 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2976-AS
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile, deionized water.
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| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Calculators
Background: BACE-1
References
- Vassar, R. et al. (1999) Science 286:735.
- Yan, R. et al. (1999) Nature 402:533.
- Cai, H. et al. (2001) Nature Neurosci. 4:233.
- Roberds, S.L. et al. (2001) Human Mol. Genet. 97:1317.
- Rockenstein, E. et al. (2005) J. Biol. Chem. 280:32957.
- Li, Q and T.C. Sudhof (2004) J. Biol. Chem. 279:10542.
- Lichtenthaler, S.F. et al. (2003) J. Biol. Chem. 278:48713.
- Kitazynem, S. et al. (2005) J. Biol. Chem. 280:8589.
Long Name
beta-site Ayloid Precursor Protein Cleaving Enzyme 1
Alternate Names
BACE1
Gene Symbol
BACE1
UniProt
Additional BACE-1 Products
Product Documents for Recombinant Mouse BACE-1 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse BACE-1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse BACE-1 Protein, CF
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Protocols
View specific protocols for Recombinant Mouse BACE-1 Protein, CF (2976-AS):
Materials
- Assay Buffer: 50 mM Sodium Acetate, 100 mM NaCl, pH 4.0
- Recombinant Mouse BACE-1 (rmBACE-1) (Catalog # 2976-AS)
- Substrate: MCA-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys(DNP)-Arg-Arg-NH2 (Catalog # ES004)
- Heparin (Sigma, Catalog # H3393), 20 mg/mL in deionized water
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmBACE-1 to 20 ng/µL in Assay Buffer.
- Dilute Heparin to 4 ng/μL in Assay Buffer.
- Combine equal volumes diluted Heparin and rmBACE-1.
- Incubate at 37 °C for 30 minutes.
- Dilute Substrate to 20 μM in Assay Buffer.
- Load 50 µL of the rmBACE-1/heparin mixture in a plate and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
Per Well:
- rmBACE-1: 0.500 µg
- Substrate: 10 µM
- Heparin: 1 ng/μL
