Recombinant Mouse BACE-1 Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Mouse BACE-1 Protein, CF Summary

Details of Functionality
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.
Source
Mouse myeloma cell line, NS0-derived mouse BACE-1 protein
Thr22-Thr457, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Thr22
Protein/Peptide Type
Recombinant Enzymes
Gene
Bace1
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
50 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
68 kDa, reducing conditions
Publications
Read Publications using
2976-AS in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile, deionized water.
Assay Procedure
  • 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
  1. Dilute rmBACE-1 to 20 ng/µL in Assay Buffer.
  2. Dilute Heparin to 4 ng/μL in Assay Buffer.
  3. Combine equal volumes diluted Heparin and rmBACE-1.
  4. Incubate at 37 °C for 30 minutes.
  5. Dilute Substrate to 20 μM in Assay Buffer.
  6. 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.
  7. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  8. 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse BACE-1 Protein, CF

  • APP beta-secretase
  • ASP2
  • Aspartyl protease 2
  • BACE1
  • BACE-1
  • BACEAsp 2
  • beta-secretase 1 precursor variant 1
  • beta-secretase 1
  • beta-site amyloid beta A4 precursor protein-cleaving enzyme
  • Beta-site amyloid precursor protein cleaving enzyme 1
  • Beta-site APP cleaving enzyme 1
  • beta-site APP-cleaving enzyme 1
  • beta-site APP-cleaving enzyme
  • EC 3.4.23
  • EC 3.4.23.46
  • FLJ90568
  • HSPC104
  • KIAA1149
  • memapsin-2
  • Membrane-associated aspartic protease 2
  • transmembrane aspartic proteinase Asp2

Background

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.

  1. Vassar, R. et al. (1999) Science 286:735.
  2. Yan, R. et al. (1999) Nature 402:533.
  3. Cai, H. et al. (2001) Nature Neurosci. 4:233.
  4. Roberds, S.L. et al. (2001) Human Mol. Genet. 97:1317.
  5. Rockenstein, E. et al. (2005) J. Biol. Chem. 280:32957.
  6. Li, Q and T.C. Sudhof (2004) J. Biol. Chem. 279:10542.
  7. Lichtenthaler, S.F. et al. (2003) J. Biol. Chem. 278:48713.
  8. Kitazynem, S. et al. (2005) J. Biol. Chem. 280:8589.

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Publications for BACE-1 (2976-AS)(2)

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Bioinformatics

Gene Symbol Bace1
Uniprot