Protease Assay Service
At Creative Biolabs, our Protease Assay Service platform enables precise measurement and profiling of both secreted and intracellular proteases, with sensitivity tailored to immune-oncology contexts—supporting tool compound identification and functional studies.
Background
Proteolytic enzymes—including granzymes, cathepsins, caspases, and matrix metalloproteases—play pivotal roles in immune-mediated cytotoxicity, antigen processing, extracellular matrix remodeling, and cytokine activation. Granzymes released by cytotoxic T or NK cells induce apoptosis in target tumor cells. Cathepsins support antigen presentation through proteolysis of endosomal proteins. MMPs modulate tumor stroma to enable immune cell infiltration. Dysregulated protease activity is directly linked to ineffective immune surveillance, tumor immune evasion, and immunosuppressive microenvironments. Accurately characterizing protease kinetics and inhibitor responses is essential to uncover immune-targeted modulators in oncology research.
Featured Services at Creative Biolabs
Our Protease Assay Service offers robust biochemical and cell-based assays tailored to enzyme class, immune function, and compound screening needs:
Fluorogenic & FRET Substrate Assays
Real-time quantification of protease activity using peptides labeled with fluorophores or donor-acceptor pairs (e.g., caspase-3/7, granzyme B, cathepsin S/B).
Activity-Based Probe (ABP) Profiling
Covalent labeling of active proteases in tumor and immune cell lysates, enabling target engagement studies.
Gel Zymography
Detection of extracellular protease activity (e.g., MMPs) in tumor-immune cell co-cultures or supernatants, enabling spatial and size-discriminated analysis.
High-throughput Screening (HTS)
Miniaturization in 96- or 384-well plates with Z' > 0.7 and robust dynamic range for SAR and inhibitor screening.
Target Coverage
Includes granzyme B, caspase-3/7, cathepsins B/K/S, MMP2/9, and other proteases relevant to immune surveillance and microenvironment regulation.
Assays can be designed using purified enzymes or immune-cell extracts, and extended to multiplexed detection or orthogonal validation formats.
Workflow
Highlights
- Immune-Function-Oriented Targets – Assay panels include granzyme B for cytotoxic response, cathepsins for antigen processing, and MMPs for microenvironment remodeling
- Multiple Detection Modalities – Real-time fluorescence, ABP capture, and gel-based zymography allow cross-validation
- Superior Throughput & Sensitivity – Z' > 0.75 ensures high-quality HTS performance
- Customizable Substrate Design – Peptides derived from tumor-associated antigens or immune checkpoint domains
- Qualified Support Team – Expert proteomics and enzymology team guiding assay design and result interpretation
FAQs
Q1: Can granzyme B activity be measured directly in immune-cell lysates?
A1: Yes. We use fluorogenic substrate assays and ABP profiling to measure granzyme B activity in CTL or NK cell extracts following immune activation.
Q2: Are secreted proteases (e.g., MMPs) quantifiable in co-culture systems?
A2: Absolutely. Gel zymography and fluorogenic MMP assays reliably quantify secreted protease activity from tumor–immune cell supernatants.
Q3: Can you discriminate between protease isoforms or family members?
A3: Yes. Substrate specificity and probe selection allow selective measurement of caspases, cathepsins, or MMP subtypes.
Q4: Do you offer kinetic parameter analysis (e.g., Vmax, Km)?
A4: Yes. We provide full enzyme kinetics including Km, Vmax, IC₅₀, and dose–response curves suitable for SAR modeling.
Q5: Is the service compatible with screening libraries or mechanistic profiling?
A5: Yes. Both focused inhibitor panels and high-throughput compound libraries are supported under validated HTS conditions.
Leverage Protease Insights for Tumor Immunology Projects
Creative Biolabs offers dedicated support through assay design, immune-contextual optimization, and data analysis services specifically focused on protease biology within oncology landscapes. Whether designing assays targeting cytotoxic granzymes or matrix-remodeling MMPs, we provide rapid consultation, custom workflows, and high-integrity reporting.
Download our brochure