详细说明
- Purity>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
- Endotoxin Level<1.0 EU per 1 μg of the protein by the LAL method.
- ActivityMeasured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH 2 (Catalog # ). The specific activity is >50 pmol/min/µg, as measured under the described conditions. See Activity Assay Protocol on www.RnDSystems.com
- SourceMouse myeloma cell line, NS0-derived Phe29-Pro462 (Tyr357His) with a C-terminal 6-His tag
- Accession #
- N-terminal Sequence
AnalysisPhe29, Ala63, Asp140 - Predicted Molecular Mass48 kDa
- SDS-PAGE55-76 kDa, reducing conditions
| 2977-AS | | |
| Formulation Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. | ||
| 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.
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- Assay Buffer: 50 mM NaOAc, 1.0 M NaCl, 0.05% Brij-35 (w/v), pH 3.0
- Recombinant Mouse BACE-2 (rmBACE-2) (Catalog # 2977-AS)
- Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # )
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmBACE-2 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 2 ng/µL rmBACE-2 in 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.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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).
- rmBACE-2: 0.1 µg
- Substrate: 10 µM
BACE-2 is an aspartic protease that shares sequence similarity with BACE-1, the major beta ‑secretase for generation of A beta peptide in neurons (1). However, BACE-2 differs from BACE-1 in several aspects including pro-enzyme activation, substrate binding sites, transcriptional regulation, and function in A beta peptide production (2‑4). The domain structure of mouse BACE-2 consists of a signal peptide (aa 1‑19), pro (aa 20‑62) and a mature chain (aa 63‑514) that consists of a transmembrane domain and a cytoplasmic tail. The C‑terminal region (aa 463‑514) was replaced by a His tag in the Recombinant Mouse BACE-2 that also contained a His residue at the position of Tyr357. The purified rmBACE-2 started at Ala63, the same residue shown at the N-terminus of the human active BACE-2 (2).
- 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. (2005) FASEB J. 19:739.
- 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
400-6226-992