ADAMTS-20 (Recombinant)
- Expression system
- HEK293
- Cat. #
- REC-ADAMTS20
In stock
- SKU
- REC-ADAMTS20
Target Overview
ADAMTS-20 (A Disintegrin and Metalloproteinase with Thrombospondin Motifs 20; UniProt P59510) is a 1,910-amino acid secreted metalloprotease belonging to the ADAMTS family of zinc-dependent endopeptidases. The full-length protein is classified under EC 3.4.24.- and is localised to the extracellular space and extracellular matrix, where it participates in tissue-remodelling processes under both homeostatic and pathological conditions. Beyond its catalytic role, ADAMTS-20 carries a GON domain implicated in a protease-independent function: facilitating transport of secretory cargo from the endoplasmic reticulum to the Golgi apparatus, making it structurally and functionally distinct from shorter ADAMTS family members. This recombinant form is produced in HEK293 mammalian cells, providing the post-translational modifications — including glycosylation and propeptide processing — that are characteristic of the native secreted protein and that are typically absent from bacterial or insect-cell expression systems. Mammalian expression is particularly relevant for ADAMTS-20 given its complex multi-domain architecture and known secretory pathway biology. Researchers use this recombinant in substrate cleavage assays, degradomic workflows designed to identify physiological substrates, and metalloprotease inhibitor IC50 determinations. It also serves as a well-defined positive control antigen for Western blot and immunohistochemistry antibody validation experiments. Laboratories working on extracellular matrix biology, pigmentation genetics, or musculoskeletal research have used recombinant ADAMTS-20 to interrogate protease–substrate relationships and to benchmark detection reagents. Researchers requiring an antibody for parallel immunodetection experiments can pair this recombinant with the Triple Point Biologics matched anti-ADAMTS-20 antibody (SKU: RP-ADAMTS20), which is validated for Western blot against human targets with predicted cross-reactivity to rat and monkey.
Background
Applications
- Substrate cleavage assay using MT1-MMP or extracellular matrix protein substrates
- Metalloprotease inhibitor IC50 determination in fluorogenic or gel-based formats
- Degradomic substrate identification by quantitative mass spectrometry (TAILS or similar N-terminomics)
- Western blot positive control for anti-ADAMTS-20 antibody validation (pairs with TPB RP-ADAMTS20)
- Immunohistochemistry antigen standard for anti-ADAMTS-20 antibody titration and specificity testing
- ELISA capture or detection standard for ADAMTS-20 quantification in conditioned media or tissue lysates
- Biophysical characterisation of ADAMTS-20 domain interactions (e.g., SPR, BLI, or pull-down with GON domain binding partners)
- Melanoblast migration and neural crest ECM remodelling in vitro assays requiring exogenous recombinant protease
References
- Lee KAV et al. Genome-Wide Association Study and Rare Variant Association Studies of Strabismus in the All of Us Research Program. Ophthalmol Sci. 2025. doi:10.1016/j.xops.2025.100873. PMID: 40837069.
- Dai D et al. Genomic analysis reveals the association of KIT and MITF variants with the white spotting in swamp buffaloes. BMC Genomics. 2024. doi:10.1186/s12864-024-10634-2. PMID: 39048931.
- El-Sayed A et al. Doppler ultrasonographic scan, gene expression and serum profile of immune, APPs and antioxidant markers in Egyptian buffalo-cows with clinical endometritis. Sci Rep. 2024. doi:10.1038/s41598-024-56258-0. PMID: 38459095.
- Nandadasa S et al. Degradomic Identification of Membrane Type 1-Matrix Metalloproteinase as an ADAMTS9 and ADAMTS20 Substrate. Mol Cell Proteomics. 2023. doi:10.1016/j.mcpro.2023.100566. PMID: 37169079.
- Cilia C et al. Whole Genome Sequencing Unravels New Genetic Determinants of Early-Onset Familial Osteoporosis and Low BMD in Malta. Genes (Basel). 2022. doi:10.3390/genes13020204. PMID: 35205249.
Additional Specifications
| Storage Buffer | 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM CaCl₂, 10% glycerol |
|---|---|
| Endotoxin Level | <0.1 EU/µg by LAL |
| Purity (%) | >90% by SDS-PAGE |
| Expression System | HEK293 |
| Subcellular Localization | Secreted; Extracellular matrix |
Frequently Asked Questions
What is the expected molecular weight of recombinant ADAMTS-20 on SDS-PAGE or Western blot?
The full-length ADAMTS-20 polypeptide (UniProt P59510) is 1,910 amino acids with a predicted unglycosylated MW of ~217 kDa. Because this recombinant is expressed in HEK293 cells, N-linked and O-linked glycosylation adds significant mass; expect a diffuse band in the 230–260 kDa range under reducing SDS-PAGE conditions. The broad appearance is characteristic of heavily glycosylated secreted proteases and is not indicative of degradation. Running alongside a high-range marker (250 kDa or above) is recommended. Our lot-specific CoA includes an annotated SDS-PAGE image for direct comparison.
What processed or secreted form of ADAMTS-20 does this recombinant represent — is the propeptide removed?
ADAMTS-20 is synthesised as a zymogen; furin-type proprotein convertases cleave the propeptide in the trans-Golgi network to yield the mature, active metalloprotease. HEK293 expression recapitulates this processing, so the protein in this vial is predominantly the propeptide-cleaved, secreted form. A minor unprocessed pro-form may co-purify, visible as a faint higher-MW band on SDS-PAGE. If full processing is critical for your assay, the lot-specific CoA documents the mature-to-pro ratio observed under our standard purification conditions.
What substrates does ADAMTS-20 cleave and which is best for a fluorometric activity assay?
ADAMTS-20 is a versican-specific aggrecanase: it cleaves versican at the Glu441–Ala442 bond within the CS-α domain of the core protein, with documented activity against versican V0 and V1 isoforms. For fluorometric assays, FRET-based synthetic peptides spanning this cleavage site (e.g., Mca-EPENFF↓EALEK-Dap(Dnp)) serve as tractable substrates. Alternatively, full-length recombinant versican incubated with ADAMTS-20 can be read out by Western blot using neo-epitope antibodies against the DPEAAE neoepitope. The latter approach is more physiologically relevant but less amenable to high-throughput formats.
What buffer and divalent cation conditions should I use for ADAMTS-20 activity assays?
ADAMTS-20 is a zinc-dependent metalloprotease that requires Ca²⁺ for full catalytic activity. The protein is supplied in 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10% glycerol, 5 mM CaCl₂ — a formulation compatible with most in vitro proteolytic assays. For activity assays, maintain at least 1–5 mM CaCl₂ in the reaction buffer; dropping below 0.5 mM CaCl₂ typically reduces activity substantially. Include 0.05% Brij-35 or BSA (0.1 mg/mL) if working at sub-nanomolar enzyme concentrations to minimise adsorption losses. EDTA or EGTA at ≥5 mM will abolish activity and can serve as negative controls.
What starting concentration of recombinant ADAMTS-20 should I use for a versican cleavage assay?
A reasonable starting point for an in vitro versican cleavage assay is 10–50 nM recombinant ADAMTS-20 with 100–500 nM recombinant versican substrate, incubated at 37 °C in 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM CaCl₂, 0.05% Brij-35 for 2–16 hours. Titrate enzyme down to determine the linear range. For FRET-peptide substrates, 1–10 nM enzyme with 10–50 µM peptide typically gives a readable initial velocity. Always include a heat-inactivated enzyme control (95 °C, 10 min) to confirm that signal is enzyme-dependent.
Can I use recombinant ADAMTS-20 as a Western blot positive control with the RP-ADAMTS20 antibody?
Yes — this is the primary intended cross-application. The matched rabbit polyclonal antibody (SKU: RP-ADAMTS20; /anti-adamts-20-rabbit-polyclonal-antibody) was validated using this recombinant as the reference antigen, so band identity and migration are confirmed. Load 20–50 ng of recombinant ADAMTS-20 per lane alongside your cell or tissue lysate. Expect a band at approximately 230–260 kDa under reducing conditions. Running a titration series (10, 25, 50 ng) in flanking lanes provides a clean signal gradient for antibody sensitivity benchmarking in your specific transfer and blocking conditions.
How much recombinant ADAMTS-20 should I load as a positive control to validate RP-ADAMTS20?
A clean single band at 230–260 kDa in the recombinant lane, absent in a secondary-only control, confirms antibody specificity before you commit tissue sections. Note that IHC signal intensity in native tissue will depend on fixation and antigen retrieval; the recombinant WB lane serves as a specificity reference, not a direct quantitative standard for IHC.
How should I handle, dilute, and store recombinant ADAMTS-20 to preserve activity over time?
The protein is supplied in 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10% glycerol, 5 mM CaCl₂ and is stable at -20 °C in single-use aliquots for up to 12 months from receipt. Avoid repeated freeze-thaw cycles — each cycle can reduce recoverable activity by 10–20% for metalloproteinases. For working dilutions, use the supplied buffer or your assay buffer supplemented with 0.1 mg/mL BSA to minimise surface adsorption, especially below 10 nM. Do not dilute into buffers containing chelators (EDTA, EGTA). Thaw on ice and use within 4 hours; do not re-freeze diluted aliquots.
Validation imagery coming soon
Western blot validation figures for REC-ADAMTS20 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.