Recombinant Human BACE-2 Protein, CF Summary
Phe29-Pro466, with a C-terminal 10-His tag
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.
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.
|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.
- 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
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).
- rhBACE-2: 0.1 µg
- Substrate: 25 µM
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).
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.
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.
- 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.
No product specific FAQs exist for this product, however you mayView all Proteins and Enzyme FAQs
Fluorogenic Peptide Substrates
Reviews for Recombinant Human BACE-2 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human BACE-2 Protein, CF and earn rewards!
Have you used Recombinant Human BACE-2 Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image