Recombinant Human BACE-2 Protein, CF

Catalog #: 4097-ASB Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
4097-ASB-020

Recombinant Human BACE-2 Protein Enzyme Activity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human BACE-2 Protein, CF Summary

Product Specifications

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 #

Q9Y5Z0-1

N-terminal Sequence
Analysis

Phe29 & Ala63

Predicted Molecular Mass

49 kDa

SDS-PAGE

48-60 kDa, reducing conditions

Product Datasheets

4097-ASB

Product Datasheet

COA

SDS

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 V_max* (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

Scientific Data

Enzyme Activity

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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

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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.

Reconstitution Calculator

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

Cai, H. et al. (2001) Nature Neurosci. 4:233.
Hussain, I. et al. (2001) J. Biol. Chem. 276:23322.
Ostermann, N. et al. (2006) J. Mol. Biol. 355:249.
Sun, X. et al. (2006) FASEB J. 19:739.
Esterhazy. D. et al. (2011) Cell Metab. 14:365.
Casas, S. et al. (2010). Am. J. Physiol. Endocrinol. Metab. 299:E1087.
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