Recombinant Human BACE-2 Protein, CF

R&D Systems | Catalog # 4097-ASB

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human BACE-2 Protein (4097-ASB)
  • 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

Q9Y5Z0-1

Applications

Enzyme Activity
View all BACE-2 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human BACE-2 protein
Phe29-Pro466, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Phe29 & Ala63

Predicted Molecular Mass

49 kDa

SDS-PAGE

48-60 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2 (Catalog # ES010).
The specific activity is >70 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human BACE-2 Protein, CF

Recombinant Human BACE-2 Protein Enzyme Activity

Recombinant Human BACE-2 Protein Enzyme Activity

Recombinant Human BACE-2 (Catalog # 4097-ASB) is measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2 (Catalog # ES010).

Recombinant Human BACE-2 Protein SDS-PAGE

Recombinant Human BACE-2 Protein SDS-PAGE

2 μg/lane of Recombinant Human BACE‑2 was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band at approximately 55 kDa under R conditions.

Formulation, Preparation, and Storage

4097-ASB
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. 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 opening.

Background: BACE-2

BACE-2 (Beta secretase 2) is an aspartic protease that shares 48% sequence identity with BACE-1 in the extracellular catalytic domains. BACE-1 is the putative beta secretase for the generation of the A beta peptide in neurons (1). BACE-2 differs from BACE-1 in several aspects, including proenzyme activation, substrate preference, transcriptional regulation, and expression pattern (2, 3). Unlike BACE-1, BACE-2 activity does not contribute to Alzheimer's disease pathogenesis (4) but has been shown to play a key role in insulin receptor trafficking in the pancreas where it is expressed in beta -cells (5,6). BACE-2 affects glucose tolerance and was suggested as a promising target for improving beta -cell function in diabetes (7). Recombinant human BACE-2 was expressed without its C terminal transmembrane and cytosolic domains, resulting in its secretion from NS0 cells.

References

  1. Cai, H. et al. (2001) Nature Neurosci. 4:233.
  2. Hussain, I. et al. (2001) J. Biol. Chem. 276:23322.
  3. Ostermann, N. et al. (2006) J. Mol. Biol. 355:249.
  4. Sun, X. et al. (2006) FASEB J. 19:739.
  5. Esterhazy. D. et al. (2011) Cell Metab. 14:365.
  6. Casas, S. et al. (2010). Am. J. Physiol. Endocrinol. Metab. 299:E1087.
  7. Alcarraz-Vizan, G. et al. (2017). Cell Mol. Life Sci. 74:2827.

Long Name

beta-site Ayloid Precursor Protein Cleaving Enzyme 2

Alternate Names

BACE2

Entrez Gene IDs

25825 (Human); 56175 (Mouse); 288227 (Rat)

Gene Symbol

BACE2

UniProt

Q9Y5Z0-1

Additional BACE-2 Products

Product Documents for Recombinant Human BACE-2 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-2 Protein, CF

For research use only

Related Research Areas

Aspartic Proteases and Regulators

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Protocols

View specific protocols for Recombinant Human BACE-2 Protein, CF (4097-ASB):

Materials
  • Assay Buffer: 50 mM Sodium Acetate, 1 M NaCl, 0.05% Brij-35, pH 3.0
  • Recombinant Human BACE-2 (rhBACE-2) (Catalog # 4097-ASB)
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES010)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1.  Dilute rhBACE-2 to 2 ng/µL in Assay Buffer.
2.  Dilute Substrate to 50 µM in Assay Buffer.
3.  Load 50 µL of 2 ng/µL rhBACE-2 in a plate, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 50 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-2: 0.1 µg
  • Substrate: 25 µM

FAQs

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