Protease Assays
Our protease assay panel provides a platform for profiling and screening new protease inhibitors against all major classes of proteases, such as serine proteases, cysteine proteases, threonine proteases, aspartic proteases, glutamic proteases, and metalloproteases.
In the human genome, there are around 500 putative proteases. Proteases play a role in physiological processes by managing complicated proteolytic activities. Proteases are targeted successfully for the treatment of virus infections such as HIV and Hepatitis C. Inhibitors have been approved for the treatment of hypertension (ACE inhibitors), diabetes (DPP-IV inhibitor), and to prevent blood clots (Factor Xa inhibitors). Studies are underway for the treatment of Alzheimer's, COVID-19, and cancer.
Reaction Biology has developed the largest protease assay panel for protease screening to ensure the most comprehensive selectivity data. Our protease assays are based on the cleavage of fluorogenic peptide substrates for quantification of the proteolytic activity of proteases enabling the screening of direct and allosteric inhibitors.
Protease panel profiling is a valuable tool to determine the selectivity of a new protease inhibitor and detect off-target effects supporting the creation of safe drug candidates.
- Fluorescence-based protease activity assay
- Low and large-scale protease screening, as well as high-throughput screening options, are available
- Full protease panel protease profiling is performed on a monthly basis
- Discover our new COVID-19-related protease assays
List of Protease Assays (76 total)
| Name | HGNC Symbol | Additional Synonyms | Data |
|---|---|---|---|
| ACE1 | ACE | Angiotensin-converting enzyme | Data sheet |
| ACE2 | ACE2 | Angiotensin-converting enzyme 2 | Data sheet |
| Activated Protein C | PROC | Vitamin K-dependent Protein C, APC, PROC | Data sheet |
| ADAM10 | ADAM10 | a disintegrin and metalloproteinase domain 10, MADM HsT18717 CD156C | Data sheet |
| BACE1 | BACE1 | Beta-secretase 1 | Data sheet |
| Calpain1 | CPN1 | CAN1, CAPN1, CANPL1 | Data sheet |
| Caspase1 | CASP1 | Interleukin-1 beta convertase, p45, IL1BC, IL1BCE | Data sheet |
| Caspase10 | CASP10 | ICE-like apoptotic protease 4, Apoptotic protease Mch-4, FAS-associated death domain protein interleukin-1B-converting enzyme 2, FLICE2 | Data sheet |
| Caspase11 | n.a. | mouse Caspase 4, Protease ICH-3, CASP4, Casp11, Caspl, Ich3 | Data sheet |
| Caspase14 | CASP14 | CASPE | Data sheet |
| Caspase2 | CASP2 | Neural precursor cell expressed developmentally down-regulated protein 2, Protease ICH-1, ICH1, NEDD2 | Data sheet |
| Caspase3 | CASP3 | Apopain, Cysteine protease CPP32, Protein Yama, SREBP cleavage activity 1, CPP32 | Data sheet |
| Caspase4 | CASP4 | ICE(rel)-II, Protease ICH-2, Protease TX, ICH2 | Data sheet |
| Caspase5 | CASP5 | ICE(rel)-III, Protease ICH-3, Protease TY, ICH3 | Data sheet |
| Caspase6 | CASP6 | Apoptotic protease Mch-2, MCH2 | Data sheet |
| Caspase7 | CASP7 | Apoptotic protease Mch-3, CMH-1, ICE-like apoptotic protease 3, MCH3 | Data sheet |
| Caspase8 | CASP8 | Apoptotic cysteine protease, Apoptotic protease Mch-5, CAP4, FADD-homologous ICE/ced-3-like protease, FADD-like ICE, ICE-like apoptotic protease 5, MORT1-associated ced-3 homolog, MCH5 | Data sheet |
| Caspase9 | CASP9 | Apoptotic protease Mch-6, Apoptotic protease-activating factor 3, ICE-like apoptotic protease 6, MCH6 | Data sheet |
| Cathepsin B | CTSB | APP secretase, Cathepsin B1, CATB, CPSB | Data sheet |
| Cathepsin C | CTSC | DPP1, PLS, PALS | Data sheet |
| Cathepsin D | CTSD | CPSD, CATD | Data sheet |
| Cathepsin E | CTSE | CATE | Data sheet |
| Cathepsin G | CTSG | CG | Data sheet |
| Cathepsin H | CTSH | CPSB, ACC-4, ACC-5, ACC4, ACC5 | Data sheet |
| Cathepsin K | CTSK | Cathepsin O/O2/X | Data sheet |
| Cathepsin L | CTSL | CTSL1, MEP | Data sheet |
| Cathepsin S | CTSS | CTSS, CATS | Data sheet |
| Cathepsin V | CTSV | CTSL2, cathepsin L2 | Data sheet |
| Chymase | CMA1 | CMA1, CYH, CYM | Data sheet |
| Chymotrypsin | n.a. | Alpha-chymotrypsin | Data sheet |
| Dipeptidyl peptidase IV (DDP-IV) | DPP4 | ADCP2, CD26 | Data sheet |
| Dipeptidyl peptidase IX (DPPIX) | DPP9 | DPRP2 | Data sheet |
| Dipeptidyl peptidase VIII (DPPVIII) | DPP8 | DPRP1 | Data sheet |
| Elastase | ELANE | Bone marrow serine protease, Elastase-2, Human leukocyte elastase, Medullasin, ELA2 | Data sheet |
| Factor IXa | F9 | Coagulation factor IX | Data sheet |
| Factor VIIa | F7 | Coagulation factor VII | Data sheet |
| Factor Xa | F10 | Coagulation factor X | Data sheet |
| Factor XIa | F11 | Coagulation factor XI, FXI | Data sheet |
| Factor XIIa | F12 | Coagulation factor XII | Data sheet |
| Furin | FURIN | FUR, PACE, PCSK3 | Data sheet |
| Granzyme B | GZMB | Granzyme-2, C11, CTLA-1 | Data sheet |
| Hepsin* | HPN | Transmembrane protease serine 1, TMPRSS1, HPN | Data sheet |
| HIV-1 | n.a. | Human immunodeficiency virus-1 | Data sheet |
| Kallikrein 1 | KLK1 | Tissue kallikrein, KLK1 | Data sheet |
| Kallikrein 12 | KLK12 | kallikrein related peptidase 12, KLK-L5 | Data sheet |
| Kallikrein 13 | KLK13 | kallikrein related peptidase 13, KLK-L4 | Data sheet |
| Kallikrein 14 | KLK14 | kallikrein related peptidase 14, KLK-L6 | Data sheet |
| Kallikrein 2 | KLK2 | kallikrein related peptidase 2, prostatic | Data sheet |
| Kallikrein 5 | KLK5 | kallikrein related peptidase 5, SCTE, KLK-L2 | Data sheet |
| Kallikrein 7 | KLK7 | kallikrein related peptidase 7, SCCE | Data sheet |
| Matriptase 2 | TMPRSS6 | Transmembrane protease serine 6 | Data sheet |
| MMP 1 | MMP1 | Matrix metalloproteinase-2, MMP-2, Gelatinase A, TBE-1 | Data sheet |
| MMP 10 | MMP10 | matrix metallopeptidase 10, STMY2 | Data sheet |
| MMP 12 | MMP12 | HME, Matrix metalloproteinase-12, MMP-12, Macrophage elastase, ME | Data sheet |
| MMP 13 | MMP13 | Matrix metalloproteinase-13, MMP-13 | Data sheet |
| MMP 14 | MMP14 | MMP-14, Membrane-type matrix metalloproteinase 1, MT-MMP 1, MTMMP1, Membrane-type-1 matrix metalloproteinase, MT1-MMP, MT1MMP, MMP-X1 | Data sheet |
| MMP 2 | MMP2 | Matrix metalloproteinase-2, MMP-2, Gelatinase A, TBE-1 | Data sheet |
| MMP 3 | MMP3 | Matrix metalloproteinase-3, MMP-3, Transin-1, SL-1 | Data sheet |
| MMP 7 | MMP7 | Pump-1 protease, Uterine metalloproteinase, Matrix metalloproteinase-7, MMP-7, Matrin | Data sheet |
| MMP 8 | MMP8 | matrix metallopeptidase 8, CLG1 | Data sheet |
| MMP 9 | MMP9 | MMP-9, 92 kDa type IV collagenase, 92 kDa gelatinase, Gelatinase B, GELB | Data sheet |
| Neprilysin | MME | matrix metalloendopeptidase, CALLA, CD10, NEP | Data sheet |
| Papain | n.a. | Papaya proteinase I | Data sheet |
| Plasma Kallikrein | KLKB1 | Fletcher factor, kallikrein B1 | Data sheet |
| Proteinase A | n.a. | n.a. | Data sheet |
| Proteinase K | n.a. | Endopeptidase K | Data sheet |
| SARS-CoV-2 Mpro | n.a. | SARS-CoV-2 Main Protease, SARS-CoV-2 3C-like Proteinase, 3CLpro | Data sheet |
| SARS-CoV-2 PLpro | n.a. | SARS-CoV-2 Papain-like Protease, NS3 | Data sheet |
| TACE | ADAM17 | A disintegrin and metalloproteinase domain 17, TNF-alpha-converting enzyme, TNF-alpha convertase, Snake venom-like protease, CD156b antigen | Data sheet |
| Thrombin | F2 | coagulation factor II | Please inquire |
| TMPRSS11D | TMPRSS11D | Human airway trypsin-like protease, HAT | Data sheet |
| TMPRSS2 | TMPRSS2 | Transmembrane protease serine 2, Serine protease 10, PRSS10, TMPS2 | Data sheet |
| tPA* | PLAT | Tissue-type plasminogen activator, Alteplase, Reteplase | Data sheet |
| Trypsin | PRSS1 | Trypsin-1 | Data sheet |
| Tryptase g1 | TPSG1 | TPSG1, Serine protease 31, Transmembrane tryptase | Data sheet |
| Urokinase | PLAU | u-plasminogen activator, upa | Please inquire |
*Protease targets not part of the monthly full screening panels
Protease Assay Details
Assay principle with fluorogenic peptide substrates. Internally quenched fluorescent peptide substrates contain a fluorophore (5-FAM) and a quencher (QXL 520). A fluorescence signal is generated upon peptide cleavage which is quantified for the determination of protease activity.
Assay principle with FRET-quencher peptide substrates. The peptide substrates contain a fluorophore and quencher on the substrate in close proximity. Energy is transferred from the fluorescent donor to the quenching acceptor. The fluorescent signal increases with the activity of the protease cleaving the substrate.
Setups: Single-dose screening in duplicates or IC50 value determination with 5 or 10 concentrations. Other screening formats are available upon request.
Controls: No inhibitor (DMSO vehicle) control and for every assay, one target-specific control compound is tested in 10-dose IC50 format.
Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.
Report: The raw data, % enzyme activity and control compound IC50 values will be reported in Excel format for single-dose assays. For IC50 orders, raw data, IC50 values, and curve fitting will be delivered in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.
Screening facility: This assay is performed at our screening facility in Malvern, PA, US.
Compound requirements: In brief, for a standard project, 20ul of a 10mM DMSO stock or solid material is needed. Less material is needed for large scale screening. Please refer to our FAQs for information regarding compound preparation and shipping.
On a monthly basis, Reaction Biology performs compound screening on the full protease panel.
Please see the upcoming screening dates here.
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