Recombinant Human BACE-2 Protein, CF

Catalog # Availability Size / Price Qty
4097-ASB-020
BACE-2 Protein Enzyme Activity
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Recombinant Human BACE-2 Protein, CF Summary

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.
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.
Source
Mouse myeloma cell line, NS0-derived human BACE-2 protein
Phe29-Pro466, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Phe29 & Ala63
Predicted Molecular Mass
49 kDa
SDS-PAGE
48-60 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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

Data Images

Enzyme Activity Enzyme Activity View Larger

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).

SDS-PAGE SDS-PAGE View Larger

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

Reconstitution Calculator

Reconstitution Calculator

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-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
Entrez Gene IDs
25825 (Human); 56175 (Mouse); 288227 (Rat)
Alternate Names
AEPLC; Asp 1; ASP1; ASP21; Aspartic-like protease 56 kDa; Aspartyl protease 1; BACE2; BACE-2; beta secretase 2; beta-secretase 2; beta-site amyloid beta A4 precursor protein-cleaving enzyme 2; Beta-site amyloid precursor protein cleaving enzyme 2; Beta-site APP cleaving enzyme 2; beta-site APP-cleaving enzyme 2; beta-site APP-cleaving enzyme 2, EC 3.4.2310ALP5656 kDa aspartic-like protease; CEAP1; Down region aspartic protease; Down syndrome region aspartic protease; DRAPBAE2; EC 3.4.23; EC 3.4.23.45; memapsin-1; Membrane-associated aspartic protease 1; theta-secretase; transmembrane aspartic proteinase Asp1

FAQs

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Fluorogenic Peptide Substrates

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