Proxiplate 384 f plus

Manufactured by PerkinElmer

Sourced in United States

The ProxiPlate-384 F Plus is a microplate designed for high-throughput screening applications. It features 384 wells and is made of a black polystyrene material. The plate is optimized for fluorescence-based assays.

Automatically generated - may contain errors

Lab products found in correlation

Ubiquitin rhodamine 110, by R&D Systems (16 mentions) Pherastar, by BMG Labtech (16 mentions) L glutathione reduced gsh, by Merck Group (15 mentions) Fitc ga, by Enzo Life Sciences (2 mentions) Synergy 4 hybrid microplate reader, by Agilent Technologies (2 mentions) Pst 60 hl plus thermo shaker, by Biosan (2 mentions) Envision plate reader, by PerkinElmer (2 mentions) Envision multimode plate reader, by PerkinElmer (1 mentions) Synergy 4, by Agilent Technologies (1 mentions) Synergy h1 hybrid, by Agilent Technologies (1 mentions) Spark plate reader, by Tecan (1 mentions) Spark multimode plate reader, by Tecan (1 mentions) Spr 102, by StressMarq Biosciences (1 mentions) Ub amc, by R&D Systems (1 mentions) Spectramax m5e, by Molecular Devices (1 mentions)

Spelling variants of this product

ProxiPlate (53 citations) ProxiPlate-384 Plus (15 citations) ProxiPlate Plus (4 citations) ProxiPlate-384 Plus F 384-shallow well microplate (2 citations)

30 protocols using proxiplate 384 f plus

Quantitative Assay for USP7 Enzyme Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 27

Each assay was performed in a final volume of 15 μL in assay buffer containing 20 mM Tris-HCl (pH 8.0, (1M Tris-HCl, pH 8.0 solution; Corning 46-031-CM)), 1 mM GSH (L-Glutathione reduced; Sigma #G4251), 0.03% BGG (0.22 μM filtered, Sigma, #G7516-25G), and 0.01% Triton X-100 (Sigma, #T9284-10L). Nanoliter quantities of either an 8-point or 10-point, 3-fold serial dilution in DMSO was pre-dispensed into assay plates (Perkin Elmer, ProxiPlate-384 F Plus, #6008269) for a final test concentration range of either 25 M to 11 nM or 25 μM to 1.3 nM, respectively. The final concentration of the enzyme (USP7, construct USP7 (208-1102) 6*His, Viva Biotech) in the assay was 62.5 μM. Final substrate (Ub-Rh110; Ubiquitin-Rhodamine 110, R&D Systems #U-555) concentration was 25 nM with [Ub-Rh110]<<Km. 5 μL of 2× enzyme was added to assay plates (pre-stamped with compound) preincubated with USP7 for 30 minutes and then 5 μL of 2×Ub-Rh110 was added to assay plates. Plates were incubated stacked for 20 minutes at room temperature before 5 μL of stop solution (final concentration of 10 mM citric acid in assay buffer (Sigma, #251275-500G)). Fluorescence was read on the Envision (Excitation at 485 nm and Emission at 535 nm; Perkin Elmer) or on the PheraSTAR (Excitation at 485 nm and Emission at 535 nm; BMG Labtech).

US10351571B2. Pyrrolotriazinones and imidazotriazinones as ubiquitin-specific protease 7 inhibitors (2019-07-16). FORMA Therapeutics, Inc. [US]. Inventors: Stephanos Ioannidis [US], Adam Charles Talbot [US], Bruce Follows [US], Alexandre Joseph Buckmelter [US], Minghua Wang [US], Ann-Marie Campbell [US].

+ Open protocol

+ Expand

Fluorescence-based Assay for USP7 Inhibitor Screening

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 9

Each assay was performed in a final volume of 15 μL in assay buffer containing 20 mM Tris-HCl (pH 8.0, (1M Tris-HCl, pH 8.0 solution; Corning 46-031-CM)), 1 mM GSH (L-Glutathione reduced; Sigma #G4251), 0.03% BGG (0.22 μM filtered, Sigma, #G7516-25G), and 0.01% Triton X-100 (Sigma, #T9284-10L). Nanoliter quantities of either an 8-point or 10-point, 3-fold serial dilution in DMSO was pre-dispensed into assay plates (Perkin Elmer, ProxiPlate-384 F Plus, #6008269) for a final test concentration range of either 25 μM to 11 nM or 25 μM to 1.3 nM, respectively. The final concentration of the enzyme (USP7, construct USP7 (208-1102) 6*His, Viva Biotech) in the assay was 62.5 pM. Final substrate (Ub-Rh110; Ubiquitin-Rhodamine 110, R&D Systems #U-555) concentration was 25 nM with [Ub-Rh110]<<Km. 5 μL of 2× enzyme was added to assay plates (pre-stamped with compound) preincubated with USP7 for 30 min and then 5 μL of 2×Ub-Rh110 was added to assay plates. Plates were incubated stacked for 20 min at room temperature before 5 μL of stop solution (final concentration of 10 mM citric acid in assay buffer (Sigma, #251275-500G)). Fluorescence was read on the Envision (Excitation at 485 nm and Emission at 535 nm; Perkin Elmer) or on the PheraSTAR (Excitation at 485 nm and Emission at 535 nm; BMG Labtech).

US10519130B2. Quinazolinones and azaquinazolinones as ubiquitin-specific protease 7 inhibitors (2019-12-31). FORMA Therapeutics, Inc. [US]. Inventors: Stephanos Ioannidis [US], Adam Charles Talbot [US], Bruce Follows [US], Alexandre Joseph Buckmelter [US], Minghua Wang [US], Ann-Marie Campbell [US], David R. Lancia, Jr. [US].

+ Open protocol

+ Expand

Fluorescence Polarization Assay for JAK Kinase Inhibitors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescence polarization measurements were performed in black 384-well plates (ProxiPlate-384 F Plus, PerkinElmer) at sample volume of 5 μl/well on PerkinElmer Envision plate reader using 480 nm excitation and 535 nm emission filters. Fluorescent tracer, Bodipy FL labeled JNJ-7706621 [compound 5 in (35 (link))] was used at 1.5 nM concentration. Recombinant JAK1 JH2 553-836-His, JAK2 JH2 503-827-His, JAK3 JH2 511-790-His, and TYK2 JH2 564-876-His proteins were used at concentrations (2 nM JAK1, 150 nM JAK2, 1 μM JAK3, 20 nM TYK2) dependent on protein-tracer dissociation constants. The assays were performed in a buffer consisting of 20 mM Tris-HCl pH 8.0, 150 mM NaCl, 20% glycerol, 0.01% Brij-35, and 2 mM DTT. ATP was titrated at concentration range of 5 nM−5 μM and fluorescence polarization values obtained were fitted against log[ATP] in GraphPad Prism to yield IC50 values. Assays were performed in triplicate.

Raivola J., Hammarén H.M., Virtanen A.T., Bulleeraz V., Ward A.C, & Silvennoinen O. (2018). Hyperactivation of Oncogenic JAK3 Mutants Depend on ATP Binding to the Pseudokinase Domain. Frontiers in Oncology, 8, 560.

+ Open protocol

+ Expand

Characterization of USP7 Enzyme Inhibition

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 125

Each assay was performed in a final volume of 15 μL in assay buffer containing 20 mM Tris-HCl (pH 8.0, (1M Tris-HCl, pH 8.0 solution; Corning 46-031-CM)), 1 mM GSH (L-Glutathione reduced; Sigma #G4251), 0.03% BGG (0.22 μM filtered, Sigma, #G7516-25G), and 0.01% Triton X-100 (Sigma, #T9284-10L). Nanoliter quantities of either an 8-point or 10-point, 3-fold serial dilution in DMSO was pre-dispensed into assay plates (Perkin Elmer, ProxiPlate-384 F Plus, #6008269) for a final test concentration range of either 25 μM to 11 nM or 25 μM to 1.3 nM, respectively. The final concentration of the enzyme (USP7, construct USP7 (208-1102) 6*His, Viva Biotech) in the assay was 62.5 pM. Final substrate (13b-Rh110, Ubiquitin-Rhodamine 110, R&D Systems #U-555) concentration was 25 nM with [Ub-Rh110]<<Km 5 μL of 2× enzyme was added to assay plates (pre-stamped with compound) preincubated with USP7 for 30 minutes and then 5 μL of 2×Ub-Rh110 was added to assay plates. Plates were incubated stacked for 20 minutes at room temperature before 5 μL of stop solution (final concentration of 10 mM citric acid in assay buffer (Sigma, #251275-500G)). Fluorescence was read on the Envision (Excitation at 485 nm and Emission at 535 nm; Perkin Elmer) or on the PheraSTAR (Excitation at 485 nm and Emission at 535 nm; BMG Labtech).

US10934299B2. Pyrrolo and pyrazolopyrimidines as ubiquitin-specific protease 7 inhibitors (2021-03-02). Valo Early Discovery, Inc. [US]. Inventors: Stephanos Ioannidis [US], Adam Charles Talbot [US], Bruce Follows [US], Alexandre Joseph Buckmelter [US], Minghua Wang [US], Ann-Marie Campbell [US], Darby Rye Schmidt [US], David Joseph Guerin [US], Justin Andrew Caravella [US], R. Bruce Diebold [US], Anna Ericsson [US], David R. Lancia, Jr. [US].

+ Open protocol

+ Expand

Fluorescence Polarization Assay for IL17A

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescence polarization (FP) assays were performed in black shallow 384-well micro plates (ProxiPlate – 384 F Plus, PerkinElmer) using Envision multimode plate reader (Perkin-Elmer). Polarization was measured by using an excitation filter of 485 nm and an emission filter of 535 nm. Each well was flashed 50 times, and the average values were used. Briefly, 5 µL of IL17A (20 nM) diluted in the assay buffer (50 mM Tris pH 8.0, 0.001% Tween) was added to compound containing wells and incubated for 60 min. To this, 5 µL of the FP probe (10 nM) was added and the reaction mixture was incubated for 120 min at room temperature prior to measuring FP. All inhibition experiments were normalized against DMSO control containing no inhibitor.

Goedken E.R., Argiriadi M.A., Dietrich J.D., Petros A.M., Krishnan N., Panchal S.C., Qiu W., Wu H., Zhu H., Adams A.M., Bodelle P.M., Goguen L., Richardson P.L., Slivka P.F., Srikumaran M., Upadhyay A.K., Wu B., Judge R.A., Vasudevan A., Gopalakrishnan S.M., Cox P.B., Stoll V.S, & Sun C. (2022). Identification and structure-based drug design of cell-active inhibitors of interleukin 17A at a novel C-terminal site. Scientific Reports, 12, 14561.

+ Open protocol

+ Expand

Hsp90 Inhibitor Binding Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Displacement studies were performed with recombinant Hsp90β(SPR-102C, StressMarq) and the Hsp90 inhibitors geldanamycin and AUY922 in an established fluorescence polarization assay.^{17 (link)} Reaction mix was created containing 60 nM Hsp90β, 5 nM FITC-GA (Enzo Life Sciences), and fluorescence polarization (FP) assay buffer (20 mM HEPES pH 7.3, 50 mM KCl, 0.01% NP-40), aliquoted into a 96-well plate and preincubated for 3 h at room temperature. Compounds were serial diluted in FP assay buffer and added to the reaction plate. DMSO concentration was normalized to 2% (v/v) in all reactions. Twenty-five microliters of reaction mixtures was transferred to shallow 384-well, black microplates (ProxiPlate-384 F plus, Perkin-Elmer, Waltham, MA) and incubated for 16 h. Endpoint FP as mP values were recorded using Synergy 4 (BioTek, Winooski, VT). The measured mP values were plotted against competitor concentration.

Thomas F.M., Goode K.M., Rajwa B., Bieberich A.A., Avramova L.V., Hazbun T.R, & Jo Davisson V. (2017). A Chemogenomic Screening Platform Used to Identify Chemotypes Perturbing HSP90 Pathways. SLAS discovery : advancing life sciences R & D, 22(6), 706-719.

+ Open protocol

+ Expand

Fluorescence Polarization Assay for Polκ-RIR Interaction

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Rev1-CT/FAM-Polκ-RIR complex diluted in phosphate buffered saline (10 μL, 0.2 μM) was added to a black 384-well plate (ProxiPlate-384 F Plus, PerkinElmer). Unlabeled Polκ-RIR (10 μL, varying concentrations) was added to give final concentrations of Polκ-RIR (0.1–1000 nM) in a total volume of 20 μL/well. The mixture was incubated at room temperature for 1 hour with gentle mixing (see Supplemental Figure 2). Fluorescence polarization was measured on a Synergy H1 Hybrid multi-mode microplate reader (Biotek, excitation 485 nM, emission 528 nM). For the pilot screen, compounds were diluted to 20 μM in PBS from a 2 mM DMSO stock solution for screening at a final concentration of 10 μM. Compounds (10 μL) and Rev1-CT/FAM-Polκ-RIR complex (10 μL, 0.2 μM) were mixed in 384-well black plates on a Microlab Nimbus (Hamilton Robotics) automated liquid handler. The mixture was incubated and analyzed as described above. All graphs and statistical analysis were carried out with GraphPad Prism 5.

Sail V., Rizzo A.A., Chatterjee N., Dash R.C., Ozen Z., Walker G.C., Korzhnev D.M, & Hadden M.K. (2017). Identification of small molecule translesion synthesis inhibitors that target the Rev1-CT/RIR protein-protein interaction. ACS chemical biology, 12(7), 1903-1912.

+ Open protocol

+ Expand

Enzymatic Assay for USP7 Inhibitor Screening

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 9

Each assay was performed in a final volume of 15 μL in assay buffer containing 20 mM Tris-HCl (pH 8.0, (1M Tris-HCl, pH 8.0 solution; Corning 46-031-CM)), 1 mM GSH (L-Glutathione reduced; Sigma #G4251), 0.03% BGG (0.22 μM filtered, Sigma, #G7516-25G), and 0.01% Triton X-100 (Sigma, #T9284-10L). Nanoliter quantities of either an 8-point or 10-point, 3-fold serial dilution in DMSO was pre-dispensed into assay plates (Perkin Elmer, ProxiPlate-384 F Plus, #6008269) for a final test concentration range of either 25 μM to 11 nM or 25 μM to 1.3 nM, respectively. The final concentration of the enzyme (USP7, construct USP7 (208-1102) 6*His, Viva Biotech) in the assay was 62.5 pM. Final substrate (Ub-Rh110; Ubiquitin-Rhodamine 110, R&D Systems #U-555) concentration was 25 nM with [Ub-Rh110]<<Km. 5 μL of 2× enzyme was added to assay plates (pre-stamped with compound) preincubated with USP7 for 30 min and then 5 μL of 2× Ub-Rh110 was added to assay plates. Plates were incubated stacked for 20 min at room temperature before 5 μL of stop solution (final concentration of 10 mM citric acid in assay buffer (Sigma, #251275-500G)). Fluorescence was read on the Envision (Excitation at 485 nm and Emission at 535 nm; Perkin Elmer) or on the PheraSTAR (Excitation at 485 nm and Emission at 535 nm; BMG Labtech).

US10519129B2. Quinazolinones and azaquinazolinones as ubiquitin-specific protease 7 inhibitors (2019-12-31). FORMA Therapeutics, Inc. [US]. Inventors: Stephanos Ioannidis [US], Adam Charles Talbot [US], Bruce Follows [US], Alexandre Joseph Buckmelter [US], Minghua Wang [US], Ann-Marie Campbell [US], David R. Lancia, Jr. [US].

+ Open protocol

+ Expand

Optimized USP7 Enzyme Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 125

Each assay was performed in a final volume of 15 μL in assay buffer containing 20 mM Tris-HCl (pH 8.0, (1M Tris-HCl, pH 8.0 solution; Corning 46-031-CM)), 1 mM GSH (L-Glutathione reduced; Sigma #G4251), 0.03% BGG (0.22 M filtered, Sigma, #G7516-25G), and 0.01% Triton X-100 (Sigma, #T9284-10L). Nanoliter quantities of either an 8-point or 10-point, 3-fold serial dilution in DMSO was pre-dispensed into assay plates (Perkin Elmer, ProxiPlate-384 F Plus, #6008269) for a final test concentration range of either 25 μM to 11 nM or 25 μM to 1.3 nM, respectively. The final concentration of the enzyme (USP7, construct USP7 (208-1102) 6*His, Viva Biotech) in the assay was 62.5 pM. Final substrate (Ub-Rh110; Ubiquitin-Rhodamine 110, R&D Systems #U-555) concentration was 25 nM with [Ub-Rh110]<<Km. 5 μL of 2× enzyme was added to assay plates (pre-stamped with compound) preincubated with USP7 for 30 minutes and then 5 μL of 2×Ub-Rh110 was added to assay plates. Plates were incubated stacked for 20 minutes at room temperature before 5 μL of stop solution (final concentration of 10 mM citric acid in assay buffer (Sigma, #251275-500G)). Fluorescence was read on the Envision (Excitation at 485 nm and Emission at 535 nm; Perkin Elmer) or on the PheraSTAR (Excitation at 485 nm and Emission at 535 nm; BMG Labtech).

US09902728B2. Pyrrolo and pyrazolopyrimidines as ubiquitin-specific protease 7 inhibitors (2018-02-27). Forma Therapeutics, Inc. [US]. Inventors: Stephanos Ioannidis [US], Adam Charles Talbot [US], Bruce Follows [US], Alexandre Joseph Buckmelter [US], Minghua Wang [US], Ann-Marie Campbell [US], Darby Rye Schmidt [US], David Joseph Guerin [US], Justin A. Caravella [US], R. Bruce Diebold [US], Anna Ericsson [US], David R. Lancia, Jr. [US].

+ Open protocol

+ Expand

Hsp90 ATPase Activity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Transcreener™ ADP kit (Bellbrook Labs) was used to assay for ATPase activity. Conditions for the Hsp90 ATPase reaction were 50 mM HEPES (pH 7.4), 20 mM KCl, 4 mM MgCl₂, and 0.01% NP-40 in a 10-μL total assay volume in shallow 384 well, black microplates (ProxiPlate- 384 F plus, Perkin Elmer) as previously described [47 (link)]. Compounds were incubated in the presence of 250 μM ATP and 0.5 μM Hsp90β at 37 °C for 30 min. ATPase activity was stopped via addition of 1X stop & detect buffer (200 mM HEPES (pH 7.5), 400 mM EDTA, and 0.2% Brij-35) with 100 mg/mL ADP² Antibody-IRDye® QC-1 and 8 nM ADP Alexa594 Tracer and incubated for 1 h at room temperature. Endpoint fluorescence intensity (RFU) was measured using a Synergy 4 hybrid microplate reader (BioTek). Experiments were performed three times and the measured RFU values were plotted against competitor concentration using GraphPad Prism and curves were fit to Sigmoidal, four parameter logistic, where X is log(concentration).

Goode K.M., Petrov D.P., Vickman R.E., Crist S.A., Pascuzzi P.E., Ratliff T.L., Davisson V.J, & Hazbun T.R. (2017). Targeting the Hsp90 C-terminal domain to induce allosteric inhibition and selective client downregulation. Biochimica et biophysica acta. General subjects, 1861(8), 1992-2006.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!