Recombinant Human BACE-1 Protein, CF

R&D Systems | Catalog # 931-AS

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human BACE-1 Protein (931-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

P56817.1

Structure / Form

Pro and Mature forms

Applications

Enzyme Activity
View all BACE-1 Proteins and Enzymes »

Product Specifications

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

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Thr22 & Glu46

Predicted Molecular Mass

50 kDa (Pro form) & 47 kDa (Mature form)

SDS-PAGE

68-70 kDa and 65-67 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 >3.5 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

931-AS
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution

Reconstitute at 500 μg/mL in sterile, deionized water.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: BACE-1

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.

References

  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.

Long Name

beta-site Ayloid Precursor Protein Cleaving Enzyme 1

Alternate Names

BACE1

Entrez Gene IDs

23621 (Human); 23821 (Mouse)

Gene Symbol

BACE1

UniProt

P56817.1

Additional BACE-1 Products

Product Documents for Recombinant Human BACE-1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human BACE-1 Protein, CF

For research use only

Related Research Areas

Aspartic Proteases and Regulators

Citations for Recombinant Human BACE-1 Protein, CF

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Protocols

View specific protocols for Recombinant Human BACE-1 Protein, CF (931-AS):

Materials
  • 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

FAQs

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