BACE-2 (Recombinant)
- Expression system
- HEK293
- Cat. #
- REC-BACE2
In stock
- SKU
- REC-BACE2
Target Overview
BACE-2 (Beta-secretase 2; UniProt Q9Y5Z0) is a type I transmembrane aspartic protease of 518 amino acids, classified under EC 3.4.23.45. It is a member of the pepsin family of aspartic endoproteases and is expressed at the cell membrane, where it mediates ectodomain shedding of several physiologically important substrates. This recombinant is produced in HEK293 cells, an expression system that supports the glycosylation patterns and folding environments characteristic of the native human enzyme — relevant for researchers who require post-translational modifications to be preserved for accurate enzymatic activity measurements or protein–protein interaction studies. BACE-2 cleaves the amyloid precursor protein (APP) at two defined sites — between residues 690/691 and 671/672 — generating soluble APP ectodomains and membrane-tethered C-terminal fragments. It also cleaves PMEL within the M-beta fragment during early melanosome biogenesis, releasing the amyloidogenic luminal fragment required for fibril assembly. Additionally, BACE-2 has been shown to process CLTRN in pancreatic beta cells, placing it at the intersection of neurobiological, pigmentation, and metabolic research contexts. In a research setting, this recombinant is used primarily in three areas: (1) enzymatic activity assays using fluorogenic or peptide substrates to characterise cleavage kinetics; (2) inhibitor screening and IC50 determination, often in parallel with BACE-1 to establish selectivity profiles; and (3) antibody validation, where the recombinant serves as a positive control antigen. Researchers performing Western blot or immunohistochemistry validation of anti-BACE-2 antibodies can pair this recombinant directly with the Triple Point Biologics matched antibody (SKU: RP-BACE2), which has been validated for Western blot against human BACE-2. Immunohistochemistry (IHC) and immunofluorescence (IF) validation is currently in progress.
Background
Applications
- Fluorogenic peptide substrate cleavage assay to measure BACE-2 aspartic protease activity (kcat/Km determination)
- Inhibitor IC50 screening and BACE-1 vs. BACE-2 selectivity profiling in parallel enzyme panels
- APP beta-site and theta-site cleavage characterisation by LC-MS/MS or gel-based fragment analysis
- PMEL M-beta fragment processing assay to study luminal amyloidogenic fragment release in melanosome biogenesis research
- Positive control antigen for Western blot validation of anti-BACE-2 antibodies (pairs with TPB RP-BACE2)
- Immunoassay calibration standard for quantification of BACE-2 in serum or conditioned media (e.g., ELISA development)
- Substrate identification and active-site mapping by hydrogen-deuterium exchange mass spectrometry (HDX-MS) or crosslinking MS
- Recombinant antigen for immunogen confirmation and epitope competition assays
References
- Li B et al. Elevated circulating BACE2 captures chronic glycemic burden and enhances the clinical identification of type 2 diabetes. iScience. 2026. doi:10.1016/j.isci.2026.115988. PMID: 42221820.
- Garvert L et al. Effects of BACE2 polymorphisms on verbal memory in the general population. Eur Arch Psychiatry Clin Neurosci. 2026. doi:10.1007/s00406-026-02208-y. PMID: 41902949.
- Polerà N et al. Upregulation of ACHE and BACE2 genes by oleacein in Alzheimer's disease and neuroblastoma. Chem Biol Interact. 2026. doi:10.1016/j.cbi.2026.111909. PMID: 41520719.
- Gandhi V et al. In Silico Investigation of Amidine-Based BACE-1 Inhibitors Against Alzheimer's Disease: SAR, Pharmacokinetics, Molecular Docking and Dynamic Simulations. Pharmaceuticals (Basel). 2025. doi:10.3390/ph19010005. PMID: 41599607.
- Liu Q et al. Integrated network toxicology and experimental validation to investigate potential mechanisms associated with aspartame-induced malignant phenotypic changes in colorectal cancer. Naunyn Schmiedebergs Arch Pharmacol. 2026. doi:10.1007/s00210-026-05591-4. PMID: 42319418.
Additional Specifications
| Storage Buffer | 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10% glycerol |
|---|---|
| Endotoxin Level | <0.1 EU/µg by LAL |
| Purity (%) | >90% by SDS-PAGE |
| Expression System | HEK293 |
| Subcellular Localization | Type I transmembrane; Golgi apparatus → endosomes → cell surface |
Frequently Asked Questions
What molecular weight should I expect for recombinant BACE-2 on SDS-PAGE or Western blot?
The full-length BACE-2 protein (UniProt Q9Y5Z0) has a calculated molecular weight of ~57 kDa from its 518-amino-acid sequence. However, because this recombinant is expressed in HEK293 cells, N-linked glycosylation adds appreciable mass — expect a diffuse band running at approximately 60–70 kDa under denaturing, reducing conditions. The smearing is characteristic of heterogeneous glycosylation and is not a purity artifact; purity is confirmed at >95% by SDS-PAGE. If you require a sharper band for quantitation, PNGase F treatment collapses the glycoforms to the ~57 kDa core.
What processing form of BACE-2 does this recombinant represent — pro-enzyme or mature enzyme?
This recombinant corresponds to the mature, processed form of BACE-2. The inhibitory prodomain (approximately the first 22 residues of the canonical sequence after signal peptide cleavage) is removed, yielding an active enzyme. The construct retains the catalytic aspartyl dyad (Asp93 and Asp289) and is delivered as a soluble, active fragment — the transmembrane anchor is absent, making it directly suitable for solution-phase activity assays without detergent solubilization steps.
What substrates does BACE-2 cleave, and what cleavage sites should I design my activity assay around?
BACE-2 cleaves amyloid precursor protein (APP) at two defined sites: between residues 690/691 (the theta-site) and 671/672 (the beta-site), generating soluble APP ectodomains and C-terminal membrane-tethered fragments. It also cleaves PMEL within the M-beta fragment during early melanosome biogenesis. For FRET-based activity assays, a peptide substrate spanning the APP theta-site (e.g., SEVNL↓DAEF, where ↓ denotes cleavage) is a well-validated choice. BACE-2 activity is inhibited by pepstatin A; including a pepstatin A control at 1–10 µM confirms on-target protease activity in your assay.
What buffer conditions and pH are optimal for a BACE-2 activity assay with this recombinant?
BACE-2 is an aspartic protease with optimal activity in mildly acidic conditions, consistent with its endosomal/cell-surface biology. For in vitro activity assays, use 50 mM sodium acetate pH 4.5–5.0 with 0.1% Triton X-100 and 0.1 mg/mL BSA as a carrier. The storage buffer for this recombinant is 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10% glycerol — dilute the enzyme at least 1:10 into your assay buffer to avoid pH carry-over from the storage buffer, which would suppress activity. Run a pH titration between 4.0 and 6.0 if you are optimizing a novel substrate.
What starting concentration of recombinant BACE-2 should I use in an enzyme activity assay?
A starting point of 5–20 nM recombinant BACE-2 in the assay well works well with FRET peptide substrates at 10–50 µM. Titrate enzyme concentration to achieve linear signal over your intended time window (typically 30–60 min at 37°C). For inhibitor IC50 determinations, keep the enzyme concentration at or below the anticipated Ki of your compound to avoid tight-binding artifacts — use the Morrison equation if [E] approaches [I]. Confirm linearity by running a 2-fold dilution series (e.g., 2.5, 5, 10, 20 nM) before committing to a full inhibitor panel.
Can I use recombinant BACE-2 as a positive control for Western blot with the matched TPB anti-BACE-2 antibody (RP-BACE2)?
Yes — this is a validated pairing. REC-BACE2 and RP-BACE2 are produced and cross-validated in the same laboratory. Load 50–150 ng of REC-BACE2 per lane alongside your cell lysate samples; after transfer and blocking, probe with RP-BACE2 at the recommended dilution (see the RP-BACE2 product page at /anti-bace-2-rabbit-polyclonal-antibody). Expect a band at 60–70 kDa (glycosylated) in the recombinant lane. Because both reagents share the same lot-to-lot validation pipeline, the recombinant lane provides a reliable size marker and signal-intensity reference for your experimental samples.
How much recombinant BACE-2 should I load as a positive control to match endogenous BACE-2 signal in cell lysates?
Endogenous BACE-2 is typically expressed at low-to-moderate levels; in most cell lines it is substantially lower signal than BACE-1. Load a two-point bracket: 50 ng and 150 ng of REC-BACE2 per lane. The 50 ng lane approximates a moderate-expression lysate signal and helps you gauge whether your lysate band falls within the linear range of the RP-BACE2 antibody. Run a 10-lane total protein normalization alongside. If your lysate signal is well below the 50 ng recombinant band, consider immunoprecipitation prior to Western blot to enrich BACE-2.
How should I store and handle recombinant BACE-2 to preserve enzymatic activity — can I refreeze unused material?
Store at -20°C in the supplied single-use aliquots in 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10% glycerol. Repeated freeze-thaw cycles measurably reduce activity — in our stability testing, three freeze-thaw cycles result in >30% loss of specific activity. On the day of use, thaw one aliquot on ice, dilute into pre-chilled assay buffer, and keep on ice until pipetting. Do not refreeze thawed material; discard unused enzyme after the experiment. For short-term use within a single week, the protein is stable at 4°C for up to 5 days without significant activity loss.
Validation imagery coming soon
Western blot validation figures for REC-BACE2 will be published here as they are produced in-house.
If you would like to see existing validation data for this antibody before publication, request a sample copy.