Recombinant Human BACE-1 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human 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 >3.5 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human BACE-1 protein
Thr22-Tyr460 (pro) & Glu46-Tyr460 (mature), both with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Thr22 & Glu46
Structure / Form
Pro and Mature forms
Protein/Peptide Type
Recombinant Enzymes
Gene
BACE1
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
50 kDa (Pro form) & 47 kDa (Mature form).
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
68-70 kDa and 65-67 kDa, reducing conditions
Publications
Read Publications using
931-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 PBS.
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Reconstitution Instructions
Reconstitute at 500 μg/mL in sterile, deionized water.
Assay Procedure
  • Assay Buffer: 0.1 M Sodium Acetate, pH 4.0
  • Recombinant Human BACE -1 (rhBACE-1) (Catalog # 931-AS)
  • Fluorogenic Peptide Substrate IV: MCA-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys(DPN)-Arg-Arg-NH2 (Catalog # ES004)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhBACE-1 to 20 ng/µL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. Load 50 µL of rhBACE-1 into 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.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. 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:
  • rhBACE-1: 1 µg
  • Substrate: 10 µM

Notes

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

Alternate Names for Recombinant Human 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 (1-5). BACE-1 is the peptidase predominantly responsible for cleavage of the amyloid precursor protein beta site in the brain to generate the amyloid beta peptide. Because the amyloid beta peptide is a major component of amyloid plaques, BACE-1 has been implicated in the onset and/or progression of Alzheimer's disease. BACE-1 is expressed in a variety of human tissues. It is likely that this peptidase has functions in addition to its hydrolysis of the amyloid precursor protein. The peptidase activity of BACE-1 is optimal under mildly acidic conditions (pH 3.5-5.5), consistent with its proposed function in an acidic intracellular compartment.

  1. Ermolieff, J. et al. (2000) Biochemistry 39:12450.
  2. Lin, X. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1456.
  3. Sinha, S. et al. (1999) Nature 402:537.
  4. Vassar, R. et al. (1999) Science 286:735.
  5. Yan, R. et al. (1999) Nature 402:533.

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Publications for BACE-1 (931-AS)(4)

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Bioinformatics

Gene Symbol BACE1
Entrez
Uniprot