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Details of RareLED ID Assay_1019
Primary information
GENECTSZ
NUMBER OF ASSAY198
ASSAY IDs1253921 746598 746595 47812 50241 51216 51175 50266 48400 181373 46710 46525 1063747 1063744 1063743 744930 51174 50286 1063746 1063745 1241503 747457 673910 620709 448880 1258333 1258332 1258331 1343919 1253922 723768 723767 723752 746599 50283 1258203 1257954 1257953 1257952 746596 1344627 691905 691904 691902 691900 691899 691777 609673 609671 609670 1258204 1258149 1257951 1344626 50242 51399 691903 691901 609669 1063748 744931 704616 271493 391116 256977 51200 50270 48411 1241504 673911 620715 620710 180467 155400 46712 46711 46528 699426 699424 699423 1259214 256992 256978 256975 1241506 48215 699425 314227 51355 723769 224787 50113 242310 703432 703431 677819 677817 677816 609674 526076 312631 312630 215814 215053 210859 210858 210857 164133 157974 157973 95174 89194 52306 51518 242380 703433 691962 691906 677818 391117 312628 48209 51224 699427 256974 256
Assay Name1. Inhibition of human liver cathepsin L using Z-FR-AMC as substrate measured every 15 sec for 5 mins by fluorescence assay,2. Inhibition of recombinant cathepsin-B (unknown origin) using Z-Arg-Arg-AMC as substrate by fluorescence assay,3. Inhibition of recombinant cathepsin-K (unknown origin) using Z-Phe-Arg-AMC as substrate by fluorescence assay,4. Compound was evaluated for the inhibition of Cathepsin B,5. Inhibition of bovine cathepsin B,6. In vitro inhibitory activity against human cathepsin L, using 2 uM of Z -Leu-Arg-AMC as substrate,7. In vitro inhibitory activity against Human Cathepsin K using 2 uM of Z -Leu-Arg-AMC as substrate,8. In vitro inhibitory activity against human cathepsin B, using 2 uM of Z -Leu-Arg-AMC as substrate,9. In vitro inhibitory activity against human cathepsin K, using 2 uM of Z -Leu-Arg-AMC as substrate,10. Platelet membrane permeability.,11. Compound was evaluated for the inhibition of Cysteine protease Calpain 2,12. Inhibition of the cysteine protease human Calpain 1,13. Inhibition of recombinant human Cathepsin L using Z-Phe-Arg-aminomethylcoumarin as substrate preincubated for 30 mins followed by substrate addition,14. Inhibition of human Cathepsin B using Z-Phe-Arg-aminomethylcoumarin as substrate,15. Inhibition of human Cathepsin B using Z-Phe-Arg-aminomethylcoumarin as substrate preincubated for 30 mins followed by substrate addition,16. Inhibition of human liver Cathepsin B using Z-Arg-Arg-aminomethylcoumarin as substrate incubated for 5 mins prior to substrate addition measured for 5 mins by fluorometric analysis,17. Inhibition of Human Cathepsin K,18. Inhibition of Human Cathepsin B,19. Inhibition of human Cathepsin V using Z-Phe-Arg-aminomethylcoumarin as substrate,20. Inhibition of human Cathepsin V using Z-Phe-Arg-aminomethylcoumarin as substrate preincubated for 30 mins followed by substrate addition,21. Inhibition of human cathepsin-K using Z-Gly-Pro-Arg-AMC as substrate preincubated for 30 mins measured after 10 mins by fluorescence assay,22. Inhibition of cathepsin L (unknown origin) using Z-FR-AMC as substrate after 30 mins by fluorescence assay,23. Inhibition of human liver cathepsin L using Z-R-R-AMC as substrate preincubated with compound for 5 mins measured after 20 mins by fluorescence analysis,24. Reversible inhibition of human recombinant cathepsin B assessed as formation of fluorescent degradation product AMC using Z-Arg-Arg-AMC as substrate by Michaelis-Menten method,25. Inhibition of cathepsin Z after 10 to 15 mins by fluorescence assay,26. Biological Assay: Recombinant human procathepsin K was obtained from Enzo Life Sciences. Activation of the proenzyme was performed in 32.5 mM sodium acetate pH 3.5, EDTA 1 mM, NaCl 500 mM, human procathepsin K 5.5 uM, at room temperature. Activation times were optimized and varied between 35 and 150 minutes. Cathepsin K was preincubated with test compounds at various concentrations for 5 minutes at 25C. The assay was initiated by addition of substrate Z-Phe-Arg-aminomethylcoumarin ("Z-F-R-AMC" Bacchem) and the final assay conditions were 1.5 nM cathepsin K, 50 uM Z-F-R-AMC, 150 mM sodium acetate pH 5.5, 2.5 mM EDTA (Omnipure), 2.5 mM DTT (EMD), 0.01% BRIJ 35 (Sigma), and 4.0% DMSO (Acros). Test compounds were serially diluted with DMSO and water to include a final concentration range of 10 uM to 10 pM. The reaction was monitored fluorometrically for 5 minutes at 25C. using black 96-well Corning 3686 assay microplates with a Thermo Fluoroskan Ascent FL microplate reader.,27. Biological Assay: Human liver Cathepsin B (Calbiochem) was pre-incubated with test compound at various concentrations for 5 minutes at 37C. The assay was initiated by addition of substrate Z-Arg-Arg-aminomethylcoumarin (Z-R-R-AMC Bacchem) and the final assay conditions were 1.1 nM cathepsin B, 60 uM Z-R-R-AMC, 126 mM sodium potassium phosphate pH 6.0 (Fisher), 0.3 mM EDTA (Omnipure), 2.7 mM DTT (Omnipure), 0.004% BRIJ 35 (Sigma), and 2.0% DMSO (Acros) in a final volume of 200 uL. Test compounds were serially diluted with DMSO and 0.01% BRIJ 35 to include a final concentration range of 20 uM to 10 pM. The reaction was monitored fluorometrically for 5 minutes at 37C. using black 96-well Corning 3686 assay microplates with a Thermo Fluoroskan Ascent FL microplate reader at excitation and emission filter wavelengths of 355 nm and 460 nm, respectively.,28. Biological Assay: Human liver Cathepsin L (Sigma) was preincubated with test compounds at various concentrations for 5 minutes at 25C. The assay was initiated by addition of substrate Z-Phe-Arg-aminomethylcoumarin (Z-F-R-AMC Bacchem) and the final assay conditions were 1 nM cathepsin L, 50 uM Z-F-R-AMC, 100 mM sodium acetate pH 5.5, 1 mM EDTA (Omnipure), 3 mM DTT (EMD), 0.01% BRIJ 35 (Sigma), and 2.0% DMSO (Acros). Test compounds were serially diluted with DMSO and water to include a final concentration range of 10 uM to 10 pM. The reaction was monitored fluorometrically for 5 minutes at 25C. using black 96-well Corning 3686 assay microplates with a Thermo Fluoroskan Ascent FL microplate reader at excitation and emission filter wavelengths of 355 nm and 460 nm, respectively.,29. Cathepsin Enzyme Inhibition Assay: Enzyme activity is measured by observing the increase in fluorescence intensity caused by cleavage of a peptide substrate containing a fluorophore whose emission is quenched in the intact peptide. Assay buffer: 100 mM potassium phosphate pH 6.5, EDTA-Na 5 mM, Triton X-100 0.001%, DDT 5 mM. Enzymes (all at 1 nM): human and mouse Cathepsin S, Cat K, Cat B, Cat LSubstrate (20 uM): Z-Val-Val-Arg-AMC, except for Cat K which uses Z-Leu-Arg-AMC (both from Bachem). Z=Benzyloxycarbonyl. AMC=7-Amino-4-Methyl-Coumarin. Final volume: 100 uL. Excitation 360 nm, Emission 465 nm. Enzyme is added to the substance dilutions in 96-well microtitre plates and the reaction is started with substrate. Fluorescence emission is measured over 20 minutes, during which time a linear increase is observed in the absence of inhibitor.,30. Inhibition of human liver cathepsin B using Z-RR-AMC as substrate after 5 mins by fluorescence assay,31. Mixed inhibition of human recombinant cathepsin B endopeptidase activity using Z-Arg-Arg-AMC substrate assessed as inhibition constant for enzyme-substrate-inhibitor complex,32. Uncompetitive inhibition of human recombinant cathepsin B endopeptidase activity using Z-Arg-Arg-AMC substrate assessed as inhibition constant for enzyme-substrate-inhibitor complex,33. Competitive inhibition of human recombinant cathepsin L endopeptidase activity using Z-FR-AMC substrate expressed in Escherichia coli assessed as inhibition constant for enzyme-inhibitor complex,34. Inhibition of recombinant cathepsin-L (unknown origin) using Z-Phe-Arg-AMC as substrate by fluorescence assay,35. Inhibition of human cathepsin B,36. Enzyme Inhibition Assay: Enzyme activity is measured by observing the increase in fluorescence intensity caused by cleavage of a peptide substrate containing a fluorophore whose emission is quenched in the intact peptide.Assay buffer: 100 mM potassium phosphate pH 6.5, EDTA-Na 5 mM, Triton X-100 0.001%, DTT 5 mM.Enzymes (all at 1 nM): human and mouse Cathepsin S, Cat K, Cat B, Cat L.Substrate (20 inverted question markM): Z-Val-Val-Arg-AMC, except for Cat K which uses Z-Leu-Arg-AMC (both from Bachem).Z=Benzyloxycarbonyl.AMC=7-Amino-4-Methyl-Coumarin.DTT=dithiothreitol.Final volume: 100 inverted question markL.Excitation 360 nm, Emission 465 nm.Enzyme is added to the substance dilutions in 96-well microtitre plates and the reaction is started with substrate. Fluorescence emission is measured over 20 minutes, during which time a linear increase is observed in the absence of inhibitor.,37. Enzyme Inhibition Assay: Enzyme activity is measured by observing the increase in fluorescence intensity caused by cleavage of a peptide substrate containing a fluorophore whose emission is quenched in the intact peptide. Assay buffer: 100 mM potassium phosphate pH 6.5, EDTA-Na 5 mM, Triton X-100 0.001%, DTT 5 mM. Enzymes (all at 1 nM): human and mouse Cathepsin S, Cat K, Cat B, Cat L. Substrate (20 inverted question markM): Z-Val-Val-Arg-AMC, except for Cat K which uses Z-Leu-Arg-AMC (both from Bachem).Z=Benzyloxycarbonyl.AMC=7-Amino-4-Methyl-Coumarin. DTT=dithiothreitol. Final volume: 100 inverted question markL. Excitation 360 nm, Emission 465 nm.,38. Enzyme Inhibition Assay: Enzyme activity is measured by observing the increase in fluorescence intensity caused by cleavage of a peptide substrate containing a fluorophore whose emission is quenched in the intact peptide. Assay buffer: 100 mM potassium phosphate pH 6.5, EDTA-Na 5 mM, Triton X-100 0.001%, DTT 5 mM. Enzymes (all at 1 nM): human and mouse Cathepsin S, Cat K, Cat B, Cat L. Substrate (20 inverted question markM): Z-Val-Val-Arg-AMC, except for Cat K which uses Z-Leu-Arg-AMC (both from Bachem).Z=Benzyloxycarbonyl.AMC=7-Amino-4-Methyl-Coumarin. DTT=dithiothreitol. Final volume: 100 inverted question markL. Excitation 360 nm, Emission 465 nm.,39. Enzyme Inhibition Assay: Enzyme activity is measured by observing the increase in fluorescence intensity caused by cleavage of a peptide substrate containing a fluorophore whose emission is quenched in the intact peptide. Assay buffer: 100 mM potassium phosphate pH 6.5, EDTA-Na 5 mM, Triton X-100 0.001%, DTT 5 mM. Enzymes (all at 1 nM): human and mouse Cathepsin S, Cat K, Cat B, Cat L. Substrate (20 inverted question markM): Z-Val-Val-Arg-AMC, except for Cat K which uses Z-Leu-Arg-AMC (both from Bachem).Z=Benzyloxycarbonyl.AMC=7-Amino-4-Methyl-Coumarin. DTT=dithiothreitol. Final volume: 100 inverted question markL. Excitation 360 nm, Emission 465 nm.,40. Inhibition of recombinant cathepsin-S (unknown origin) using Z-Val-Arg-AMC as substrate by fluorescence assay,41. Enzyme Inhibition Assay: Enzyme activity is measured by observing the increase in fluorescence intensity caused by cleavage of
ACTIVITYSubstance BioActivity: 12 Active, 8 Activity <= 1 µM, 15 Tested Substance BioActivity: 3 Active, 45 Tested Substance BioActivity: 2 Active, 6 Tested Substance BioActivity: 2 Active, 2 Tested Substance BioActivity: 45 Active, 38 Activity <= 1 µM, 45 Tested Substance BioActivity: 22 Active, 5 Activity <= 1 µM, 38 Tested Substance BioActivity: 1 Active, 1 Activity <= 1 µM, 1 Tested Substance BioActivity: 17 Active, 3 Activity <= 1 µM, 38 Tested Substance BioActivity: 40 Active, 34 Activity <= 1 µM, 41 Tested Substance BioActivity: 10 Active, 32 Tested Substance BioActivity: 88 Active, 84 Activity <= 1 µM, 89 Tested Substance BioActivity: 86 Active, 72 Activity <= 1 µM, 86 Tested Substance BioActivity: 1 Active, 1 Activity <= 1 µM, 1 Tested Substance BioActivity: 1 Active, 1 Tested Substance BioActivity: 1 Active, 1 Tested Substance BioActivity: 30 Tested Substance BioActivity: 13 Active, 2 Activity <= 1 nM, 13 Activity <= 1 µM, 13 Tested Substance BioActivity: 12 Active, 6 Activ