| Assay Method Information | |
| | Enzyme Assay |
| Description: | Recombinant human cathepsins (CatS, CatK, CatL, CatB) were purchased from a Enzo Life Sciences. All assays were carried out in 96-well format using a buffer of 50 mM KH2PO4, 50 mM NaCl, 2 mM EDTA, 0.5 mM DTT and 1% Triton-X-100, pH 6.5 for Cathepsin S and a buffer of 50 mM NaOAc, 10 mM EDTA, 1 mM DTT and 0.01% Triton-X-100, pH 5.5 for CatK/L/B. For CatS, the enzyme (0.0007 mU/well) was incubated with fluorogeninc substrate (Z-VVR-AMC, 5 μM) at RT for 10 min. For CatK the enzyme (0.00175 mU/well) was incubated with fluorogeninc substrate (Z-FR-AMC, 40 μM) at RT for 10 min. For CatL, the enzyme (0.000874 mU/well) was incubated with fluorogeninc substrate (Z-VVR-AMC, 40 μM) at RT for 10 min. For CatB the enzyme was incubated with fluorogenic substrate (Z-VVR-AMC, 200 μM) at RT for 10 min. Flourogenic substrate turnover was detected using a microplate reader (Synergyl H4, BioTek). Ki values were calculated using the Cheng Prusoff equation (Cheng & Prosoff 1973). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |