ADAMTS-9 (Recombinant)
- Expression system
- HEK293
- Cat. #
- REC-ADAMTS9
In stock
- SKU
- REC-ADAMTS9
Target Overview
ADAMTS-9 (UniProt Q9P2N4; gene: ADAMTS9) is a secreted, extracellular matrix-localised zinc metalloprotease belonging to the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family. The full-length human protein spans 1,935 amino acids and carries the canonical ADAMTS domain architecture: a prodomain, metalloprotease domain, disintegrin-like domain, thrombospondin type-1 repeat, cysteine-rich domain, spacer region, and multiple additional thrombospondin repeats. This recombinant is produced in HEK293 mammalian cells, which supports the glycosylation and folding required for full enzymatic activity — an important consideration for a secreted, extracellular protease of this complexity. Bacterially expressed constructs of ADAMTS family members frequently lack the post-translational processing needed for reliable kinetic measurements. ADAMTS-9 cleaves two well-characterised substrates: aggrecan, at the Glu¹⁸³⁸–Ala¹⁸³⁹ bond, and versican, at the Glu¹⁴²⁸–Ala¹⁴²⁹ bond. Both cleavage events are detectable by neoepitope antibodies or mass spectrometry, making this recombinant directly suitable for substrate cleavage assays and kinetic characterisation. Separately from its protease activity, ADAMTS-9 has a protease-independent role in facilitating ER-to-Golgi transport of secretory cargo proteins, a function under active investigation. Researchers using this recombinant for antibody validation — for instance, confirming specificity in Western blot or IHC workflows — can pair it with the Triple Point Biologics matched antibody (SKU: RP-ADAMTS9), which has been validated for both applications. This recombinant serves as a well-defined positive-control antigen for those experiments.
Background
Applications
- Aggrecanase activity assay: cleavage of recombinant aggrecan substrate at the Glu1838–Ala1839 bond, detected by neoepitope antibody or SDS-PAGE
- Versican cleavage assay: proteolytic processing at the Glu1428–Ala1429 site, monitored by Western blot with anti-DPEAAE neoepitope antibody
- Metalloprotease inhibitor IC50 determination: kinetic dose-response profiling of small-molecule or endogenous inhibitor candidates against defined substrate
- Antibody validation positive control: use as recombinant antigen standard to confirm specificity of anti-ADAMTS-9 antibodies in Western blot and IHC (pairs with RP-ADAMTS9)
- Substrate identification by mass spectrometry: incubation with candidate ECM protein substrates followed by LC-MS/MS-based cleavage-site mapping
- Biophysical characterisation: SPR or ITC-based binding studies with ADAMTS inhibitors, pro-domain interactions, or matrix glycosaminoglycans
- ER-to-Golgi trafficking studies: use as a well-defined secretory cargo marker in cell-free or cell-based transport reconstitution assays examining the protease-independent function of ADAMTS-9
References
- Erkoç-Kaya D et al. CAN the variants in the THADA and ADAMST9 genes be common risk factors for type 2 diabetes development and obesity. Mol Biol Rep. 2026. doi:10.1007/s11033-026-12184-0. PMID: 42319597.
- Shen FF et al. ADAMTS9-AS2 acts as an epigenetic brake to constrain DNMT3B-mediated CADM2 silencing in esophageal squamous cell carcinoma metastasis. Front Immunol. 2026. doi:10.3389/fimmu.2026.1752827. PMID: 41869309.
- Sun Y et al. Endothelial Cell Differentiation-Related CircRNAs Drive the Differentiation of Vascular Endothelial Cells. Biochem Genet. 2026. doi:10.1007/s10528-026-11343-z. PMID: 41779083.
- Fischer S et al. Loss of ADAMTS9 disrupts ciliogenesis and collagen homeostasis resulting in Nephronophthisis-like polycystic kidneys. bioRxiv. 2026. doi:10.64898/2025.12.02.691883. PMID: 41542618.
- Liu J et al. ADAMTS9-AS2 Disrupts Docetaxel-Resistance in Castration-Resistant Prostate Cancer via Stemness Suppression and Ferroptosis Induction. Adv Sci (Weinh). 2026. doi:10.1002/advs.202520838. PMID: 41457913.
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 molecular weight should I expect for recombinant ADAMTS-9 on SDS-PAGE or Western blot?
The full-length human ADAMTS-9 protein (1,935 aa) has a predicted unglycosylated MW of ~213 kDa, but because this recombinant is produced in HEK293 cells it carries native-like N- and O-glycosylation. On reducing SDS-PAGE, expect the predominant band between 250–280 kDa. The exact migration will vary slightly with gel percentage; we recommend a 4–12% gradient gel. If you are running this as a positive control alongside our matched antibody RP-ADAMTS9, load the ladder lane adjacent to the recombinant lane for accurate size comparison.
Is recombinant ADAMTS-9 the full-length protein or a processed / truncated form, and does it retain the prodomain?
The recombinant is produced as the full-length precursor in HEK293 cells and undergoes intracellular furin-mediated prodomain removal during secretion, consistent with endogenous ADAMTS-9 processing. The secreted, prodomain-cleaved mature form is what is present in solution. This is important for activity assays: bacterially expressed constructs that retain the prodomain are typically latent and require separate activation steps that can introduce variability. The mature HEK293-expressed form is enzymatically active as supplied.
What substrates does recombinant ADAMTS-9 cleave, and at which specific cleavage sites?
ADAMTS-9 is an aggrecanase that cleaves two well-characterised proteoglycan substrates. For aggrecan, cleavage occurs at the Glu¹⁸³⁸–Ala¹⁸³⁹ bond within the interglobular domain. For versican, cleavage occurs at the Glu¹⁴²⁸–Ala¹⁴²⁹ bond. Both are canonical aggrecanase sites shared with ADAMTS-4 and -5, so selectivity controls matter in mixed-enzyme systems. Fluorogenic peptide substrates mimicking the aggrecan IGD sequence (e.g., Mca-YVADAPE-Dpa) or native aggrecan and versican preparations can all be used to measure ADAMTS-9 activity in vitro.
What buffer conditions are recommended for ADAMTS-9 activity assays, and does the enzyme require calcium?
ADAMTS-9 is a zinc metalloprotease and requires divalent cations for catalytic activity. The storage buffer — 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10% glycerol, 5 mM CaCl₂ — is compatible with direct use in activity assays. For assay buffer, a standard formulation of 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM CaCl₂, 0.05% Brij-35 (to reduce non-specific surface binding) works well. Avoid EDTA or EGTA; concentrations above 1 mM will chelate the active-site zinc and abolish activity. DTT above 1 mM can also reduce enzyme stability.
What starting concentration of recombinant ADAMTS-9 should I use for a fluorogenic peptide cleavage assay?
A reasonable starting point for fluorogenic substrate assays (e.g., Mca-based aggrecan peptide mimetics) is 50–200 nM recombinant ADAMTS-9 with substrate concentrations spanning 10–200 µM to bracket the expected Km. Because ADAMTS-9 is a large, multi-domain enzyme, non-specific adsorption to plate surfaces is a practical concern at lower concentrations; including 0.05% Brij-35 in the assay buffer helps. For native substrate (aggrecan or versican) cleavage assays detected by neoepitope antibody, 1–10 nM enzyme is often sufficient over a 4–18 h incubation at 37°C.
Can I use recombinant ADAMTS-9 as a positive control for Western blot with the RP-ADAMTS9 antibody?
Yes — this is a primary use case. REC-ADAMTS9 and the matched rabbit polyclonal RP-ADAMTS9 are developed in the same laboratory specifically to be compatible. Load 20–50 ng of REC-ADAMTS9 per lane on a 4–12% SDS-PAGE gel; under reducing conditions you should detect a broad band at 250–280 kDa with RP-ADAMTS9 at a 1:1,000–1:2,000 dilution. This positive control confirms antibody performance and provides a size reference when probing cell lysates or conditioned media. Because glycosylation shifts migration, using the HEK293-expressed recombinant as a control is more representative than bacterially produced fragments.
How much recombinant ADAMTS-9 should I load for Western blot when validating RP-ADAMTS9 for a new application?
Start with a two-point titration: 20 ng and 50 ng per lane. At 20 ng, RP-ADAMTS9 at 1:1,000 should give a clean signal at 250–280 kDa with minimal background on nitrocellulose or PVDF. If you are optimising for a new cell line or tissue lysate where endogenous ADAMTS-9 levels are uncertain, running a titration series (10, 25, 50 ng) alongside your lysate in the same blot lets you bracket the endogenous band directly. RP-ADAMTS9 is validated for Western blot; cross-reactivity with mouse ADAMTS-9 is predicted based on sequence homology but should be confirmed in your system.
How should I store and handle recombinant ADAMTS-9 to maintain activity over time, and what is its shelf life?
REC-ADAMTS9 is supplied in 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10% glycerol, 5 mM CaCl₂ and shipped in single-use aliquots on dry ice. Store at −20°C immediately on receipt; do not refreeze after thawing. Under these conditions, activity is stable for at least 12 months from the date of manufacture when stored properly. For working dilutions prepared on the day of assay, keep on ice and use within 4–6 hours. Adding carrier protein (e.g., 0.1% BSA) to diluted working stocks at concentrations below 10 nM can reduce adsorptive losses.
Validation imagery coming soon
Western blot validation figures for REC-ADAMTS9 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.