External ID: CHEMBL1930345

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2012
Volume: 47
First Page: 167
Last Page: 174
DOI: 10.1016/j.ejmech.2011.10.039

External ID: USP8 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP8: Primary Screen
1.#Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2.#Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3.#Reagent, 2.5 uL of USP8 in assay buffer (final concentration of 1 nM) to columns 1-46
4.#Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 50 nM) to all wells
5.#Time, Incubate at room temperature for 30 minutes
6.#Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP8 (USP8_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

External ID: CHEMBL1930346

Protocol: N/A

Comment: Compounds with activity <= 10uM or explicitly reported as active by ChEMBL are flagged as active in this PubChem assay presentation.

Journal: Eur. J. Med. Chem.
Year: 2012
Volume: 47
First Page: 167
Last Page: 174
DOI: 10.1016/j.ejmech.2011.10.039

Target ChEMBL ID: CHEMBL382
ChEMBL Target Name: CCRF-CEM
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: USP17 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP17: Primary Screen
1. Compound, 50 nL of screening compounds (final concentration of 50 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP17 in assay buffer (final concentration of 1 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
5. Time, Incubate at room temperature for 3 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP17 (USP17_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

External ID: USP7 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP7: Primary Screen
1. Reagent, 2.5 uL of USP7 in assay buffer (final concentration of 4 nM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076)
2. Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 (columns 45-46: DMSO control)
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Time, Incubate at room temperature for 60 minutes
5. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
6. Time, Incubate at room temperature for 10 minutes
7. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP7 (USP7_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

External ID: CHEMBL671529

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1993
Volume: 36
Issue: 20
First Page: 2938
Last Page: 2942
DOI: 10.1021/jm00072a013

Target ChEMBL ID: CHEMBL614149
ChEMBL Target Name: E6SM
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL3755836

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2016
Volume: 108
First Page: 616
Last Page: 622
DOI: 10.1016/j.ejmech.2015.12.029

External ID: CHEMBL3755834

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2016
Volume: 108
First Page: 616
Last Page: 622
DOI: 10.1016/j.ejmech.2015.12.029

Target ChEMBL ID: CHEMBL4338
ChEMBL Target Name: Purine nucleoside phosphorylase
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: CHEMBL3755981

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2016
Volume: 108
First Page: 616
Last Page: 622
DOI: 10.1016/j.ejmech.2015.12.029

Target ChEMBL ID: CHEMBL1250346
ChEMBL Target Name: 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

External ID: CHEMBL3755839

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2016
Volume: 108
First Page: 616
Last Page: 622
DOI: 10.1016/j.ejmech.2015.12.029

Target ChEMBL ID: CHEMBL3751658
ChEMBL Target Name: Purine nucleoside phosphorylase
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

External ID: CHEMBL3755838

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2016
Volume: 108
First Page: 616
Last Page: 622
DOI: 10.1016/j.ejmech.2015.12.029

External ID: CHEMBL3755837

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2016
Volume: 108
First Page: 616
Last Page: 622
DOI: 10.1016/j.ejmech.2015.12.029

External ID: CHEMBL3755982

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2016
Volume: 108
First Page: 616
Last Page: 622
DOI: 10.1016/j.ejmech.2015.12.029

External ID: CHEMBL695944

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1993
Volume: 36
Issue: 20
First Page: 2938
Last Page: 2942
DOI: 10.1021/jm00072a013

Target ChEMBL ID: CHEMBL399
ChEMBL Target Name: HeLa
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL1273415

Protocol: N/A

Comment: Journal: J Med Chem
Year: 2010
Volume: 53
Issue: 22
First Page: 7958
Last Page: 7966
DOI: 10.1021/jm100593s

Target ChEMBL ID: CHEMBL612462
ChEMBL Target Name: Enterovirus C
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL1273416

Protocol: N/A

Comment: Journal: J Med Chem
Year: 2010
Volume: 53
Issue: 22
First Page: 7958
Last Page: 7966
DOI: 10.1021/jm100593s

Target ChEMBL ID: CHEMBL612462
ChEMBL Target Name: Enterovirus C
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL849749

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1985
Volume: 28
Issue: 11
First Page: 1636
Last Page: 1643
DOI: 10.1021/jm00149a016

Target ChEMBL ID: CHEMBL229
ChEMBL Target Name: Alpha-1a adrenergic receptor
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

External ID: CHEMBL857218

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1993
Volume: 36
Issue: 20
First Page: 2938
Last Page: 2942
DOI: 10.1021/jm00072a013

Target ChEMBL ID: CHEMBL399
ChEMBL Target Name: HeLa
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL671181

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1993
Volume: 36
Issue: 20
First Page: 2938
Last Page: 2942
DOI: 10.1021/jm00072a013

Target ChEMBL ID: CHEMBL614149
ChEMBL Target Name: E6SM
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: USP28 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP28: Primary Screen
1. Compound, 25 nL of screening compounds (final concentration of 25 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP28 in assay buffer (final concentration of 0.5 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 12 nM) to all wells
5. Time, Incubate at room temperature for 30 minutes
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP28 (USP28_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

External ID: CHEMBL671183

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1993
Volume: 36
Issue: 20
First Page: 2938
Last Page: 2942
DOI: 10.1021/jm00072a013

Target ChEMBL ID: CHEMBL614149
ChEMBL Target Name: E6SM
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: USP10 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP10: Primary Screen
1. Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP10 in assay buffer (final concentration of 15 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 15 nM) to all wells
5. Time, Incubate at room temperature for 60 minutes
6. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP10 (USP10_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

External ID: UCHL1 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of UCHL1: Primary Screen
1. Reagent, 2.5 uL of UCHL1 in assay buffer (final concentration of 1 nM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076)
2. Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 (columns 45-46: DMSO control)
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Time, Incubate at room temperature for 60 minutes
5. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 7 nM) to all wells
6. Time, Incubate at room temperature for 10 minutes
7. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of UCHL1 (UCHL1_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

External ID: CHEMBL671185

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1993
Volume: 36
Issue: 20
First Page: 2938
Last Page: 2942
DOI: 10.1021/jm00072a013

Target ChEMBL ID: CHEMBL614149
ChEMBL Target Name: E6SM
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: OTUD3 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of OTUD3: Primary Screen
1. Compound, 25 nL of screening compounds (final concentration of 25 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of OTUD3 in assay buffer (final concentration of 12 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
5. Time, Incubate at room temperature for 2 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of OTUD3 (OTUD3_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

External ID: USP30 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP30: Primary Screen
1. Compound, 25 nL of screening compounds (final concentration of 25 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP30 in assay buffer (final concentration of 1 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 12 nM) to all wells
5. Time, Incubate at room temperature for 2 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP30 (USP30_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.